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  • Saccharomyces cerevisiae  (60)
  • evolution
  • Springer  (102)
  • American Association for the Advancement of Science
  • American Meteorological Society
  • Periodicals Archive Online (PAO)
  • 1995-1999  (102)
  • 1990-1994
  • 1935-1939
  • 1995  (102)
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  • 1995-1999  (102)
  • 1990-1994
  • 1935-1939
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  • 1
    Electronic Resource
    Electronic Resource
    Quantitative genetics of zooplankton life histories (1995)
    Springer
    Cellular and molecular life sciences 51 (1995), S. 454-464 
    Details
    ISSN: 1420-9071
    Keywords: Quantitative genetics ; life history ; evolution ; cladocera ; heritability ; Daphnia ; zooplankton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Quantitative genetic techniques are powerful tools for use in understanding the microevolutionary process. Because of their size, lifespan, and ease of culture, many zooplankton species are ideal for quantitative genetic approaches. As model systems, studies of zooplankton life histories are becoming increasingly used for examination of the central paradigms of evolutionary theory. Two of the fundamental empirical questions that zooplankton quantitative genetics studies can answer are: 1) How much genetic variance exists in natural populations for life history traits? 2) What is the empirical evidence for trade-offs that permeate life history theory based on optimality approaches? A review of existing data onDaphnia indicates substantial genetic variance for body size, clutch size, and age at first reproduction. Average broad-sense heritabilities for these three characters across 19 populations of 6 species are 0.31, 0.31, and 0.34, respectively. Although there is some discrepancy between the two pertinent studies that were designed to decompose the total genetic variance into its additive and non-additive components, a crude average seems to suggest that approximately 60% of the total genetic variance has an additive basis. The existing data are somewhat inconsistent with respect to presence/absence of trade-offs (negative genetic correlations) among life history traits. A composite of the existing data seems to argue against the existence of strong trade-offs between offspring size and offspring number, between present and future reproduction, and between developmental rate and fecundity. However, there is some evidence for a shift toward more negative (less positive) covariances in more stressful environments (e.g., low food). Zooplankton will prove to be very useful in future study in several important areas of research, including the genetics and physiology of aging, the importance of genotype-environment interaction for life history traits, and the evolution of phenotypic plasticity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02143198
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  • 2
    Electronic Resource
    Electronic Resource
    On the elevated intestinal pH of higher termites (Isoptera: Termitidae) (1995)
    Springer
    Insectes sociaux 42 (1995), S. 57-69 
    Details
    ISSN: 1420-9098
    Keywords: Hindgut ; alkalinity ; evolution ; symbionts ; gut morphology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The pH of the gut contents was measured in 52 species of higher termites (Termitidae), representing 36 genera in all four subfamilies. A statistically significant trend was shown from lower termites with low mean gut pH through to the Termitinae with higher mean gut pHs. Elevation of the pH occurred principally in the first and third proctodaeal segments, reaching values as high as 10.5 in 8 soil-feeding genera and 1 wood-feeding genus of Termitinae. Elevation of gut pH within the Termitidae appears to be independent of the general nature of the feeding substrate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01245699
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  • 3
    Electronic Resource
    Electronic Resource
    Expression of a yeast gene can be blocked by insertion of short yeast DNA fragments between a UAS and the TATA box (1995)
    Springer
    Current genetics 27 (1995), S. 387-389 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; URS ; FBP1 Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have constructed a plasmid, pOV10, which facilitates the introduction of putative upstream activating sequences (UAS) or upstream repressing sequences (URS) from yeast genes into plasmids containing CYC1-lacZ fusions. We have observed that the insertion of yeast sequences from 155 to 195 bp between the UAS and the TATA box of a CYC1-lacZ fusion gene can block β-galactosidase expression. It is suggested that this block is related to the formation of nucleosomes on the DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352109
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  • 4
    Electronic Resource
    Electronic Resource
    NCA3, a nuclear gene involved in the mitochondrial expression of subunits 6 and 8 of the Fo-F1 ATP synthase of S. cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 409-416 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial synthesis ; Nuclear control ; F1Fo-ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Respiratory-competent nuclear mutants have been isolated which presented a cryosensitive phenotype on a non-fermentative carbon source, due to a dysfunctioning of the mitochondrial F1-Fo ATP synthase which results from a relative defect in subunits 6 and 8 of the Fo sector. Both proteins are mtDNA-encoded, but the defect is due to the simultaneous presence of a mutation in two unlinked nuclear genes (NCA2 and NCA3, for Nuclear Control of ATPase) promoting a modification of the expression of the ATP8-ATP6 co-transcript (formerly denoted AAP1-OLI2). This co-transcript matures at a unique site to give two co-transcripts of 5.2 and 4.6 kb in length: in the mutant, the 5.2-kb co-transcript was greatly lowered. NCA3 was isolated from a wild-type yeast genomic library by genetic complementation. The level of the 5.2-kb transcript, like the synthesis of subunits 6 and 8, was partly restored in the transformed strain. A 1011-nucleotide ORF was identified that encodes an hydrophilic protein of 35417 Da. Disruption of chromosomal DNA within the reading frame promoted a dramatic decrease of the 5.2-kb mRNA but did not abolish the respiratory competence of a wild-type strain. NCA3 is located on chromosome IV and produces a single 1780-b transcript.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00311209
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  • 5
    Electronic Resource
    Electronic Resource
    Failure to detect an antimutator phenotype following disruption of the Saccharomyces cerevisiae DDR48 gene (1995)
    Springer
    Current genetics 27 (1995), S. 496-500 
    Details
    ISSN: 1432-0983
    Keywords: Antimutator ; DDR48 ; Saccharomyces cerevisiae ; Spontaneous mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The antimutator phenotype, reportedly conferred by disruption of the Saccharomyces cerevisiae DDR48 gene, was suggested to affect only a specific spontaneous mutational pathway. We attempted to identify the types of mutation that are DDR48-dependent by determining the specificity of the ddr48 antimutator. However, disruption of DDR48 did not decrease the rates of spontaneous forward mutation in a plasmid-borne copy of the yeast SUP4-o gene, the reversion or suppression of the lys2–1 allele, or forward mutation at the CAN1 locus. Interestingly, the latter gene had been reported previously to be subject to the antimutator effect. DNA sequence analysis of spontaneous SUP4-o mutations arising in DDR48 and ddr48 backgrounds provided no evidence for a reduction in the rates of individual mutational classes. Thus, we were unable to verify that disruption of DDR48 causes an antimutator phenotype.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00314438
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  • 6
    Electronic Resource
    Electronic Resource
    Purification and binding properties of the Mal63p activator of Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 509-516 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Maltose fermentation ; MAL63 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mal63p is a transcriptional activator for maltose fermentation in Saccharomyces cerevisiae. We have purified it to homogeneity from a yeast strain in which the MAL63 gene is under the control of the GAL1–GAL10 promoter. Purification included fractionation of a whole-cell extract by ion-exchange chromatography, chromatography using both non-specific DNA-affinity (calf thymus), and sequence-specific DNA-affinity chromatography. Mal63p activity was assayed by its binding to a fragment of the MAL61–MAL62 promoter, using both filter-binding and electrophoretic-mobility shift assays. DNase-I footprinting identified a new binding site (site 3) between the two previously known sites (sites 1 and 2). Mal63p is a dimer, and methylation-protection experiments identify the recognition motif as: c/a GC N9 c/a GC/g.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00314440
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  • 7
    Electronic Resource
    Electronic Resource
    The PSO4 gene of S. cerevisiae is important for sporulation and the meiotic DNA repair of photoactivated psoralen lesions (1995)
    Springer
    Current genetics 27 (1995), S. 207-212 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; pso4-1 mutant Sporulation ; DNA repair ; Meiotic recombination Induced mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have evaluated the effect of the Saccharomyces cerevisiae pso4-1 mutation in sporulation and DNA repair during meiosis. We have found that pso4-1 cells were arrested in an early step of meiosis, before premeiotic DNA synthesis, and hence did not produce spores. These results suggest that the PSO4 gene may act at the start point of the cell cycle, as do some SPO and CDC genes. The pso4-1 mutant cells are specifically sensitive to 8-MOP- and 3-CPs-photoinduced lesions, and are found to be severely affected in meiotic recombination as well as impaired in the mutagenic response, as previously described for mitosis. This means that the PSO4 gene is important for the repair 8-MOP-photoinduced lesions, mainly double-strand breaks, and the processing of these lesions into recombinogenic intermediates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326150
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  • 8
    Electronic Resource
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    Determination of chromosome copy numbers in Saccharomyces cerevisiae strains via integrative probe and blot hybridization techniques (1995)
    Springer
    Current genetics 27 (1995), S. 217-228 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome copy numbers ; Ploidy probes ; Industrial yeasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methods have been devised for analyzing chromosome copy numbers in S. cerevisiae strains that may be polyploid or aneuploid, as is apparent in the case of many industrial strains. The initial step involved transformation of a strain with an integrative “ploidy probe” transplacement fragment that enable the copy number of the targeted chromosomal locus to be determined via genomic Southern blotting and quantitative probe hybridization. Dual probe co-hybridization to Southern genomic DNA blots was used to extend such locus copy number determinations to other loci within the same chromosome, thereby screening for internal consistency along the length of the chromosome. This approach was also used to extend the analysis to other chromosomes in the genome. The method was established and verified with euploid series laboratory strains and then used to examine chromosome copy numbers in three industrial strains. One brewing strain apparently contained three copies of the chromosomes tested, whilst another brewing and a baking strain showed evidence of aneuploidy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326152
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  • 9
    Electronic Resource
    Electronic Resource
    Cloning and expression of an Aspergillus kawachii endo-1,4-β-xylanase gene in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 28 (1995), S. 467-473 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus kawachii ; β-xylanase ; Expression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract First-strand cDNA was prepared from mRNA isolated from Aspergillus kawachii IFO4308 and the β-xylanase gene (xynC) amplified by using the polymerase chain reaction (PCR) technique. This gene was inserted between the yeast phosphoglycerate kinase (PGK1) gene promoter (PGK1 p) and terminator (PGK1 T) sequences. The PGK1 P-xynC-PGK1 T construct (designated XYN3) was cloned into a multicopy episomal plasmid and the XYN3 gene was expressed in Saccharomyces cerevisiae. Functional β-xylanase (Xyn3) was produced and secreted by the recombinant yeast. Xyn3 was stable between 30 and 50°C, and the optimum temperature and pH were shown to be at 60°C and lower than pH3, respectively. An autoselective fur1::LEU2 XYN3 recombinant strain was developed that allowed β-xylanase production at a level of 300 nkat/ml in a non-selective complex medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00310817
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  • 10
    Electronic Resource
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    Strategy for deletion of complete open reading frames in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 306-308 
    Details
    ISSN: 1432-0983
    Keywords: Gene deletion ; Open reading frame ; Saccharomyces cerevisiae ; Polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The classical disruption method for yeast genes is by using in vitro deletion of the gene of interest, or of a part of it, with restriction enzymes. We are now routinely using a strategy that takes advantage of polymerase chain reactions (PCRs) which amplify large pieces of DNA. Since this approach results in a complete, precise deletion of the open reading frame, which is replaced by a unique restriction site, the ligated PCR can be used for the insertion of different markers of for two-step gene disruptions without an inserted marker. As we have now used this strategy for the deletion of more than ten genes we have in this report included some hints based on our experience.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352097
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  • 11
    Electronic Resource
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    Molecular cloning and characterization of a novel gene of Candida albicans, CDR1, conferring multiple resistance to drugs and antifungals (1995)
    Springer
    Current genetics 27 (1995), S. 320-329 
    Details
    ISSN: 1432-0983
    Keywords: Multidrug resistance ; Candida albicans ; Saccharomyces cerevisiae ; ABC transporters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By functional complementation of a PDR5 null mutant of Saccharomyces cervisiae, we have cloned and sequenced the multidrug-resistance gene CDR1 of Candida albicans. Transformation by CDR1 of a PDR5-disrupted host hypersensitive to cycloheximide and chloramphenicol resulted in resistance to cycloheximide, chloramphenicol and other drugs, such as the antifungal miconazole, with collateral hypersensitivity to oligomycin, nystatin and 2,4 dinitrophenol. Our results also demonstrate the presence of several PDR5 complementing genes in C. albicans, displaying multidrug-resistance patterns different from PDR5 and CDR1. The nucleotide sequence of CDR1 revealed that, like PDR5, it encodes a putative membrane pump belonging to the ABC (ATP-binding cassette) superfamily. CDR1 encodes a 1501-residue protein of 169.9 kDa whose predicted structural organization is characterized by two homologous halves, each comprising a hydrophobic region with a set of six transmembrane stretches, preceded by a hydrophilic nucleotide binding fold.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352101
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  • 12
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    The Saccharomyces cerevisiae HIS3 and LYS2 genes complement the Schizosaccharomyces pombe his5-303 and lys1-131 mutations, respectively: new selectable markers and new multi-purpose multicopy shuttle vectors, pSP3 and pSP4 (1995)
    Springer
    Current genetics 28 (1995), S. 380-383 
    Details
    ISSN: 1432-0983
    Keywords: Autonomously replicating sequence ; Auxotrophy ; Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Cloning vector ; Selectable marker ; HIS/his ; LYS/lys
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three new S. pombe plasmids are described. Plasmids pSP3 and pSP4 are two Schizosaccharomyces pombe ars1 multicopy vectors with the Saccharomyces cerevisiae HIS3 or LYS2 genes as selectable markers. They complement the S. pombe his5-303 or lys1-131 mutations, respectively. Plasmid pSPars1 is a vector carrying the S. pombe ars1 and a unique NdeI site which allows the introduction of any selectable marker therefore bringing a unified vector backbone for the construction of new S. pombe/S. cerevisiae/E. coli shuttle vectors. These plasmids permit classical molecular genetic techniques to be performed directly.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326437
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  • 13
    Electronic Resource
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    Calmodulin-dependent protein kinase II and calmodulin are required for induced thermotolerance in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 27 (1995), S. 190-193 
    Details
    ISSN: 1432-0983
    Keywords: Calmodulin ; Calmodulin-dependent protein kinase II ; Heat shock response ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We show here that yeast mutants lacking calmodulin-dependent protein kinase II fail to fully acquire induced thermotolerance. A similar result was also obtained with mutants depending solely on either the N-terminal half or the C-terminal half of calmodulin. These findings indicate that both calmodulin-dependent protein kinase II and calmodulin are required for induced thermotolerance.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313434
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  • 14
    Electronic Resource
    Electronic Resource
    Control of glycolytic gene expression in the budding yeast (Saccharomyces cerevisiae) (1995)
    Springer
    Current genetics 29 (1995), S. 1-9 
    Details
    ISSN: 1432-0983
    Keywords: Glycolysis ; Transcriptional activation ; Saccharomyces cerevisiae ; Chromatin structure ; Glucose induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313187
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  • 15
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    A mutant allele of the SUP45 (SAL4) gene of Saccharomyces cerevisiae shows temperature-dependent allosuppressor and omnipotent suppressor phenotypes (1995)
    Springer
    Current genetics 27 (1995), S. 417-426 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Omnipotent suppression ; Nonsense suppression ; SUP45
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using a plasmid-based termination-read-through assay, the sal4-2 conditional-lethal (temperature-sensitive) allele of the SUP45 (SAL4) gene was shown to enhance the efficiency of the weak ochre suppressor tRNA SUQ5 some 10-fold at 30°C. Additionally, this allele increased the suppressor efficiency of SRM2-2, a weak tRNAGln ochre suppressor, indicating that the allosuppressor phenotype is not SUQ5-specific. A sup + sal4-2 strain also showed a temperature-dependent omnipotent suppressor phenotype, enhancing readthrough of all three termination codons. Combining the sal4-2 allele with an efficient tRNA nonsense suppressor (SUP4) increased the temperature-sensitivity of that strain, indicating that enhanced nonsense suppressor levels contribute to the conditional-lethality conferred by the sal4-2 allele. However, UGA suppression levels in a sup + sal4-2 strain following a shift to the non-permissive temperature reached a maximum significantly below that exhibited by a non-temperature sensitive SUP4 suppressor strain. Enhanced nonsense suppression may not therefore be the primary cause of the conditional-lethality of this allele. These data indicate a role for Sup45p in translation termination, and possibly in an additional, as yet unidentified, cellular process.
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    URL: http://dx.doi.org/10.1007/BF00311210
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  • 16
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    Mutants of Saccharomyces cerevisiae sensitive to oxidative and osmotic stress (1995)
    Springer
    Current genetics 27 (1995), S. 427-434 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Oxidative stress ; Osmotic stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Although oxidative stress is involved in many human diseases, little is known of its molecular basis in eukaryotes. In a genetic approach, S. cerevisiae was used to identify elements involved in oxidative stress. By using hydrogen peroxide as an agent for oxidative stress, 34 mutants were identified. All mutants were recessive and fell into 16 complementation groups (pos1 to pos16 for peroxide sensitivity). They corresponded to single mutations as shown by a 2:2 segregation pattern. Enzymes reportedly involved in oxidative stress, such as glucose-6-phosphate dehydrogenase, glutathione reductase, superoxide dismutase, as well as glutathione concentrations, were investigated in wild-type and mutant-cells. One complementation group lacked glucose-6-phosphate dehydrogenase and was shown to be allelic to the glucose-6-phosphate dehydrogenase structural gene ZWF1/MET19. In other mutants all enzymes supposedly involved in oxidative-stress resistance were still present. However, several mutants showed strongly elevated levels of glutathione reductase, gluconate-6-phosphate dehydrogenase and glucose-6-phosphate dehydrogenase. One complementation group, pos9, was highly sensitive to oxidative stress and revealed the same growth phenotype as the previously described yap1/par1 mutant coding for the yeast homologue of mammalian transcriptional activator protein, c-Jun, of the proto-oncogenic AP-1 complex. However, unlike par1 mutants, which showed diminished activities of oxidative-stress enzymes and glutathion level, the pos9 mutants did not reveal any such changes. In contrast to other recombinants between pos mutations and par1, the sensitivity did not further increase in par1 pos9 recombinants, which may indicate that both mutations belong to the same regulating circuit. Interestingly, ten complementation groups were, in parallel, sensitive to osmotic stress, and one mutant allele revealed increased heat sensitivity. Our results indicate that a surprisingly large number of genes seem to be involved in oxidative-stress resistance and a possible overlap exists between osmotic stress and other stress reactions.
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    URL: http://dx.doi.org/10.1007/BF00311211
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  • 17
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    The expression of a specific 2-deoxyglucose-6P phosphatase prevents catabolite repression mediated by 2-deoxyglucose in yeast (1995)
    Springer
    Current genetics 28 (1995), S. 101-107 
    Details
    ISSN: 1432-0983
    Keywords: 2-deoxyglucose ; 2-deoxyglucose-6P phosphatase ; Catabolite repression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 2-deoxyglucose (2-DOG), a non-metabolize analogue of glucose, is taken up by yeast using the same transporter(s) as glucose and is phosphorylated by hexokinases producing 2-deoxyglucose-6-P. We found that in DOG R yeasts, 2-DOG was not able to trigger glucose repression, even at concentrations of 0.5%. This result suggests that the specific 2-DOG-6P phosphatase, the enzyme responsible for the DOG R phenotype, may be involved in inhibiting the process of catabolite repression mediated by 2-DOG
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    URL: http://dx.doi.org/10.1007/BF00315774
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  • 18
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    Carbon catabolite regulation of transcription of nuclear genes coding for mitochondrial proteins in the yeast Kluyveromyces lactis (1995)
    Springer
    Current genetics 28 (1995), S. 267-273 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Kluyveromyces lactis ; Transcriptional regulation ; Catabolite repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Promoter regions of the KlQCR7, KlQCR8 and KlCYC1 genes, coding for subunits of the bc 1-complex and cytochrome c respectively, in the shortterm Crabtree-negative yeast Kluyveromyces lactis differ markedly in sequence from their Saccharomyces cerevisiae counterparts. They have, however, conserved very similar configurations of binding-site motifs for various transcription factors known to be involved in global and carbon-source regulation in S. cerevisiae. To investigate the carbon source-dependent expression of these genes in K. lactis, we have carried out medium-shift experiments and determined transcript levels during the shifts. In sharp contrast to the situation in S. cerevisiae, the level of expression in K. lactis is not affected when glucose is added to a non-fermentable carbon-source medium. However, the genes are not constitutively expressed, but become significantly induced when the cells are shifted from glucose to a nonfermentable carbon source. Finally, induction of transcriptional activation does not occur in media containing both glucose and non-femmentable carbon sources.
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    URL: http://dx.doi.org/10.1007/BF00309786
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  • 19
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    Positive and negative elements involved in the differential regulation by heme and oxygen of the HEM13 gene (coproporphyrinogen oxidase) in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 28 (1995), S. 503-511 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; HEM13 regulation ; Heme and oxygen ; CYP1, ROX1, SSN6, TUP1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Saccharomyces cerevisiae HEM13 gene codes for coproporphyrinogen oxidase (CPO), an oxygen-requiring enzyme catalysing the sixth step of heme biosynthesis. Its transcription is increased 40–50-fold in response to oxygen- or heme-deficiency. We have analyzed CPO activity and HEM13 mRNA levels in a set of isogenic strains carrying single or double deletions of the CYP1 (HAP1), ROX1, SSN6, or TUPI genes. The cells were grown in the presence or absence of oxygen and under heme-deficiency (hem1Δ background). Both Rox1p and Cyp1p partially repressed HEM13 in aerobic heme-sufficient cells, probably in an independent manner. In the absence of heme, Cyp1p activated HEM13 and strongly repressed ROX1, allowing de-repression of HEM13. Cyp1p had no effect on HEM13 expression in anaerobic cells. Deletions of SSN6 or TUP1 dramatically de-repressed HEM13 in aerobic cells. A series of deletions in the HEM13 promoter identified at least four regulatory regions that are required for HEM13 regulation. Two regions, containing motifs similar to the Rox1p consensus sequences, act as repression sites under aerobic growth. The two other sites act as activation sequences required for full induction under oxygen- or heme-deficiency. Taken together, these results suggest that induction of HEM13 occurs in part through relief of repression exerted by Rox1p and Cyp1p, and in part by activation mediated partly by Cyp1p under heme-deficiency and by unknown factors under oxygen-deficiency.
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    URL: http://dx.doi.org/10.1007/BF00518161
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  • 20
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    Characterization of a novel α-amylase from Lipomyces kononenkoae and expression of its gene (LKA1) in Saccharomyces cerevisiae (1995)
    Springer
    Current genetics 28 (1995), S. 526-533 
    Details
    ISSN: 1432-0983
    Keywords: α-Amylase ; Lipomyces kononenkoae ; LKA1 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A highly active α-amylase (76 250 Da) secreted by the raw starch-degrading yeast Lipomyces kononenkoae strain IGC4052B was purified and characterized. Using high performance liquid chromatography (HPLC), end-product analysis indicated that the L. kononenkoae α-amylase acted by endo-hydrolysis on glucose polymers containing α-1,4 and α-1,6 bonds, producing mainly maltose, maltotriose and maltotetraose. The following NH2-terminal amino acids were determined for the purified enzyme: Asp-Cys-Thr-Thr-Val-Thr-Val-Leu-Ser-Ser-Pro-Glu-Ser-Val-Thr-Gly. The L. kononenkoae α-amylase-encoding gene (LKA1), previously cloned as a cDNA fragment, was expressed in Saccharomyces cerevisiae under the control of the PGK1 promoter. The native signal sequence efficiently directed the secretion of the glycosylated protein in S. cerevisiae. De-glycosylation of the enzyme indicated that post-translational glycosylation is different in S. cerevisiae from that in L. kononenkoae. Zymogram analysis indicated that glycosylation of the protein in S. cerevisiae had a negative effect on enzyme activity. Southern-blot analysis revealed that there is only a single LKA1 gene present in the genome of L. kononenkoae.
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    The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters (1995)
    Springer
    Archives of microbiology 163 (1995), S. 322-328 
    Details
    ISSN: 1432-072X
    Keywords: Isocitrate lyase ; n-Alkane-utilizable yeast ; Candida tropicalis ; Saccharomyces cerevisiae ; Promoters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The upstream region of the isocitrate lyase gene (UPR-ICL, 1530bp) of an n-alkane-utilizable yeast, Candida tropicalis, induced gene expression in another yeast, Saccharomyces cerevisiae, when the yeasts were grown on acetate. Surprisingly, UPR-ICL displayed the same regulatory function in the bacterium Escherichia coli when grown on acetate. We determined the interesting nucleotide sequence of UPR-ICL. The deletion analysis of UPR-ICL in both cells revealed the presence of two distinct promoters: one was localized at-394 to-379 and regulated gene expression in S. cerevisiae; the other was tocated near the initiation codon and regulated gene expression in E. coli. The two promoter sequences were similar, but not identical to regulatory elements that have been previously reported in S. cerevisiae and E. coli, respectively. Accordingly, the possibility of novel regulatory mechanisms could not be excluded. This is an interesting example of the presence of distinct cis-acting regulatory elements responsible for the induction of gene expression in one gene by acetate in both S. cerevisiae and E. coli. Preservation of such promoters through evolution is also discussed.
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    The upstream region of the isocitrate lyase gene (UPR-ICL) of Candida tropicalis induces gene expression in both Saccharomyces cerevisiae and Escherichia coli by acetate via two distinct promoters (1995)
    Springer
    Archives of microbiology 163 (1995), S. 322-328 
    Details
    ISSN: 1432-072X
    Keywords: Key words Isocitrate lyase ; n-Alkane-utilizable yeast ; Candida tropicalis ; Saccharomyces cerevisiae ; Promoters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The upstream region of the isocitrate lyase gene (UPR-ICL, 1530bp) of an n-alkane-utilizable yeast, Candida tropicalis, induced gene expression in another yeast, Saccharomyces cerevisiae, when the yeasts were grown on acetate. Surprisingly, UPR-ICL displayed the same regulatory function in the bacterium Escherichia coli when grown on acetate. We determined the interesting nucleotide sequence of UPR-ICL. The deletion analysis of UPR-ICL in both cells revealed the presence of two distinct promoters: one was localized at –394 to –379 and regulated gene expression in S. cerevisiae; the other was located near the initiation codon and regulated gene expression in E. coli. The two promoter sequences were similar, but not identical to regulatory elements that have been previously reported in S. cerevisiae and E. coli, respectively. Accordingly, the possibility of novel regulatory mechanisms could not be excluded. This is an interesting example of the presence of distinct cis-acting regulatory elements responsible for the induction of gene expression in one gene by acetate in both S. cerevisiae and E. coli. Preservation of such promoters through evolution is also discussed.
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    Fluctuation of body temperature and cuckoo brood parasitism (1995)
    Springer
    Ecological research 10 (1995), S. 321-325 
    Details
    ISSN: 1440-1703
    Keywords: body temperature ; brood parasitism ; cuckoo ; evolution ; telemetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Body temperatures of 11 bird species, including cuckoos, were measured in an artificial meteorological room. Ratios of change in body temperature to that in air temperature were thereby obtained for each species. Cuckoos demonstrate a remarkably high value, indicating a particularly low ability to regulate body temperature. Viewed in this light, the cuckoo's parasitic behavior is very likely an adaptation to overcome a physiological disadvantage. This in turn might be expected to reinforce delay in evolution of temperature homeostasis.
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    Facts, myths and legends on the prime industrial microorganism (1995)
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    Journal of industrial microbiology and biotechnology 14 (1995), S. 514-522 
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    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Molecular taxonomy ; Classification ; Alcoholic fermentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Archaic speculations and firmly established legends regarding the origin of the yeastSaccharomyces cerevisiae and related species are revisited in light of past and recent ecological evidence pointing to a strict association with artificial, man-made environments such as wineries and fermentation plants. The nomenclature within this industrially important group is also discussed in view of the modifications imposed from application of molecular techniques to classification.
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    URL: http://dx.doi.org/10.1007/BF01573967
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    Identification of a novel moth sex pheromone inEriocrania cicatricella (Zett.) (Lepidoptera: Eriocraniidae) and its phylogenetic implications (1995)
    Springer
    Journal of chemical ecology 21 (1995), S. 29-43 
    Details
    ISSN: 1573-1561
    Keywords: Eriocrania cicatricella ; Eriocrania sparrmannella ; Eriocraniidae ; Lepidoptera ; sex pheromone ; EAG ; GC-EAD ; mass spectrometry ; synthesis ; evolution ; (Z)-4-hepten-2-one ; (2R)-heptan-2-ol ; (2R)-(Z)-4-hepten-2-ol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Extracts from different body parts of adult femaleEriocrania cicatricella (Zett.) were tested for electrophysiological activity on conspecific male antennae. Extracts from the Vth abdominal segment, containing a pair of exocrine glands, elicited the largest electroantennographic response when compared to extracts of other body parts. Female extracts were analyzed by gas chromatography with simultaneous flame ionization and electroantennographic detection (EAD). The EAD active peaks were identified as (Z)-4-hepten-2-one, (2R)-heptane-2-ol, and (2R)-(Z)-4-hepten-2-ol by coinjection on a gas chromatography and by comparison of mass spectra with those of synthetic standards. In field tests, a blend of these three pheromone components was highly attractive to conspecific males, and a subtractive assay confirmed that the unsaturated alcohol is the major pheromone component, whereas no definite behavioral activity could be assigned to the ketone or the saturated alcohol. A bait containing the two alcohols withS-configuration was attractive to maleE. sparrmannella (Bosc), whereas no males ofE. cicatricella were found in these traps. The sex pheromone compounds inE. cicatricella are chemically similar to pheromones reported in Trichoptera and they are produced in homologous glands.
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    Allozymic characterization and evolutionary relationships in the BrazilianAkodon cursor species group (Rodentia-Cricetidae) (1995)
    Springer
    Biochemical genetics 33 (1995), S. 283-295 
    Details
    ISSN: 1573-4927
    Keywords: Akodon ; Cricetidae rodents ; genetic diversity ; biochemical polymorphism ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The present study involved an electrophoretic survey of 22 protein loci in 269 individuals belonging to three species of the genusAkodon, A. aff.cursor (2n=16),A. cursor (2n=14/15), andA. montensis (2n=24/25/26), collected in Eastern Brazil. The joint results of gene diversity, genetic distances, phenetic analyses, and phylogenetic trees suggested thatA. aff.cursor has recently separated fromA. cursor and that the three species have experienced a recent chromosomal divergence followed by low allozyme differentiation. These data are in agreement with their classification as sibling species.
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    Amino acid sequence of the ribosomal protein HS23 from the halophilicHaloarcula marismortui and homology studies to other ribosomal proteins (1995)
    Springer
    The protein journal 14 (1995), S. 189-195 
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    ISSN: 1573-4943
    Keywords: Ribosomal proteins ; protein sequencing ; evolution ; Haloarcula marismortui
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The ribosomal protein HS23 from the 30S subunit of the extreme halophilicHaloarcula marismortui, belonging to the group of archaea, was isolated either by RP-HLPLC or two-dimensional polyacrylamide gel electrophoresis. The complete amino acid sequence was determined by automated N-terminal microsequencing. The protein consists of 123 residues with a corresponding molecular mass of 12,552 Da as determined by electrospray mass spectroscopy; the pI is 11.04. Homology studies reveal similarities to the eukaryotic ribosomal protein S8 fromHomo sapiens, Rattus norvegicus, Leishmania major, andSaccharomyces cerevisiae.
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    Genetic and biochemical analyses of yeast RNase MRP (1995)
    Springer
    Molecular biology reports 22 (1995), S. 75-79 
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    ISSN: 1573-4978
    Keywords: ribosome synthesis ; RNA processing ; RNase MRP ; rRNA ; Saccharomyces cerevisiae ; snoRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RNase MRP cleaves the yeast pre-rRNA at a site in internal transcribed spacer 1 (ITS1) and this cleavage can be reproducedin vitro by the highly purified enzyme. Two protein components (Pop1p and Pop2p) have been identified which are common to yeast RNase MRP and RNase P. Moreover, purified RNase P can also cleave the pre-rRNA substratein vitro, underlining the similarities between these particles. Genetic evidence suggests that RNase MRP functionally interacts with the snoRNPs which are required for other pre-rRNA processing reactions.
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    Molecular cloning, characterization and expression analysis of two catalase isozyme genes in barley (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1005-1014 
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    ISSN: 1573-5028
    Keywords: evolution ; genome mapping ; isozymes ; oxygen radicals ; powdery mildew
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Clones representing two distinct barley catalase genes, Cat1 and Cat2, were found in a cDNA library prepared from seedling polysomal mRNA. Both clones were sequenced, and their deduced amino acid sequences were found to have high homology with maize and rice catalase genes. Cat1 had a 91% deduced amino acid sequence identity to CAT-1 of maize and 92% to CAT B of rice. Cat2 had 72 and 79% amino acid sequence identities to maize CAT-2 and-3 and 89% to CAT A of rice. Barley, maize or rice isozymes could be divided into two distinct groups by amino acid homologies, with one group homologous to the mitochondria-associated CAT-3 of maize and the other homologous to the maize peroxisomal/glyoxysomal CAT-1. Both barley CATs contained possible peroxisomal targeting signals, but neither had favorable mitochondrial targeting sequences. Cat1 mRNA occurred in whole endosperms (aleurones plus starchy endosperm), in isolated aleurones and in developing seeds, but Cat2 mRNA was virtually absent. Both mRNAs displayed different developmental expression patterns in scutella of germinating seeds. Cat2 mRNA predominated in etiolated seedling shoots and leaf blades. Barley genomic DNA contained two genes for Cat1 and one gene for Cat2. The Cat2 gene was mapped to the long arm of chromosome 4, 2.9 cM in telomeric orientation from the mlo locus conferring resistance to the powdery mildew fungus (Erysiphe graminis f.sp. hordei).
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    Isolation, expression, and evolution of the gene encoding mitochondrial elongation factor Tu in Arabidopsis thaliana (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1057-1070 
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    ISSN: 1573-5028
    Keywords: Arabidopsis ; EF-Tu ; evolution ; gene families ; mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized a second nuclear gene (tufM) in Arabidopsis thaliana that encodes a eubacterial-like protein synthesis elongation factor Tu (EF-Tu). This gene does not closely resemble the previously described Arabidopsis nuclear tufA gene, which encodes the plastid EF-Tu, and does not contain sequence elements found in all cyanobacterial and plastid tufA genes. However, the predicted amino acid sequence includes an N-terminal extension which resembles an organellar targeting sequence and shares three unique sequence elements with mitochondrial EF-Tu's, from Saccharomyces cerevisiae and Homo sapiens, suggesting that this gene encodes the Arabidopsis mitochondrial EF-Tu. Consistent with this interpretation, the gene is expressed at a higher level in flowers than in leaves. Phylogenetic analysis confirms the mitochondrial character of the sequence and indicates that the human, yeast, and Arabidopsis tufM genes have undergone considerably more sequence divergence than their cytoplasmic counterparts, perhaps reflecting a cross-compartmental acceleration of gene evolution for components of the mitochondrial translation apparatus. As previously observed for tufA, the tufM gene is present in one copy in Arabidopsis but in several copies in other species of crucifers.
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    Cloning of the amphibolic Calvin cycle/OPPP enzyme d-ribulose-5-phosphate 3-epimerase (EC 5.1.3.1) from spinach chloroplasts: functional and evolutionary aspects (1995)
    Springer
    Plant molecular biology 29 (1995), S. 1279-1291 
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    ISSN: 1573-5028
    Keywords: carbon fixation ; oxidative pentose phosphate pathway ; chloroplasts ; evolution ; endosymbiosis ; isoenzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Exploiting the differential expression of genes for Calvin cycle enzymes in bundle-sheath and mesophyll cells of the C4 plant Sorghum bicolor L., we isolated via subtractive hybridization a molecular probe for the Calvin cycle enzyme d-ribulose-5-phosphate 3-epimerase (R5P3E) (EC 5.1.3.1), with the help of which several full-size cDNAs were isolated from spinach. Functional identity of the encoded mature subunit was shown by R5P3E activity found in affinity-purified glutatione S-transferase fusions expressed in Escherichia coli and by three-fold increase of R5P3E activity upon induction of E. coli overexpressing the spinach subunit under the control of the bacteriophage T7 promoter, demonstrating that we have cloned the first functional ribulose-5-phosphate 3-epimerase from any eukaryotic source. The chloroplast enzyme from spinach shares about 50% amino acid identity with its homologues from the Calvin cycle operons of the autotrophic purple bacteria Alcaligenes eutrophus and Rhodospirillum rubrum. A R5P3E-related eubacterial gene family was identified which arose through ancient duplications in prokaryotic chromosomes, three R5P3E-related genes of yet unknown function have persisted to the present within the E. coli genome. A gene phylogeny reveals that spinach R5P3E is more similar to eubacterial homologues than to the yeast sequence, suggesting a eubacterial origin for this plant nuclear gene.
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    The yeast,Saccharomyces cerevisiae, RNase P/MRP ribonucleoprotein endoribonuclease family (1995)
    Springer
    Molecular biology reports 22 (1995), S. 87-93 
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    ISSN: 1573-4978
    Keywords: ribonucleoprotein endoribonuclease ; RNase MRP ; RNase P ; Saccharomyces cerevisiae ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ribonuclease P (RNase P) is a ribonucleoprotein responsible for the endonucleolytic cleavage of the 5′-termini of tRNAs. Ribonuclease MRP (RNase MRP) is a ribonucleoprotein that has the ability to cleave both mitochondrial RNA primers presumed to be involved in mitochondrial DNA replication and rRNA precursors for the production of mature rRNAs. Several lines of evidence suggest that these two ribonucleoproteins are related to each other, both functionally and evolutionarily. Both of these enzymes have activity in the nucleus and mitochondria. Each cleave their RNA substrates in a divalent cation dependent manner to generate 5′-phosphate and 3′-OH termini. In addition, the RNA subunits of both complexes can be folded into a similar secondary structure. Each can be immunoprecipitated from mammalian cells with Th antibodies. In yeast, both have been found to share at least one common protein. This review will discuss some of the recent advances in our understanding of the structure, function and evolutionary relationship of these two enzymes in the yeast,Saccharomyces cerevisiae.
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    Ribonuclease P from plant nuclei and photosynthetic organelles (1995)
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    Molecular biology reports 22 (1995), S. 139-145 
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    ISSN: 1573-4978
    Keywords: chloroplast ; cyanelle ; evolution ; pre-tRNA processing ; ribozyme ; wheat germ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RNase P consists of both protein and RNA subunits in all organisms and organelles investigated so far, with the exception of chloroplasts and plant nuclei where no enzyme-associated RNA has been detected to date. Studies on substrate specificity revealed that cleavage by plant nuclear RNase P is critically dependent on a complete and intact structure of the substrate. No clearcut answer is yet possible regarding the order of processing events at the 5′ or 3′ end of tRNAs in the case of nuclear or chloroplast processing enzymes. RNase P from a phylogenetically ancient photosynthetic organelle will be discussed in greater detail: The enzyme from theCyanophora paradoxa cyanelle is the first RNase P from a photosynthetic organelle which has been shown to contain an essential RNA subunit. This RNA is strikingly similar to its counterpart from cyanobacteria, yet it lacks catalytic activity. Properties of the holoenzyme suggest an intermediate position in RNA enzyme evolution, with an eukaryotic-type, inactive RNA and a prokaryotic-type small protein subunit. The possible presence of an RNA component in RNase P from plant nuclei and modern chloroplasts will be discussed, including a critical evaluation of some criteria that have been frequently applied to elucidate the subunit composition of RNase P from different organisms.
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    Evolution of the Group 1 late embryogenesis abundant (Lea) genes: analysis of the Lea B19 gene family in barley (1995)
    Springer
    Plant molecular biology 28 (1995), S. 1039-1054 
    Details
    ISSN: 1573-5028
    Keywords: ABA-inducible genes ; coding region repeats ; embryo-specific gene family ; evolution ; Hordeum vulgare L. ; phylogenetic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The highly conserved Group 1 late embryogenesis abundant (Lea) genes are present in the genome of most plants as a gene family. Family members are conserved along the entire coding region, especially within the extremely hydrophilic internal 20 amino acid motif, which may be repeated. Cloning of Lea Group 1 genes from barley resulted in the characterization of four family members named B19.1, B19.1b, B19.3 and B19.4 after the presence of this motif 1, 1, 3 and 4 times in each gene, respectively. We present here the results of comparative and evolutionary analyses of the barley Group 1 Lea gene family (B19). The most important findings resulting from this work are (1) the tandem clustering of B19.3 and B19.4, (2) the spatial conservation of putative regulatory elements between the four B19 gene promoters, (3) the determination of the relative ‘age’ of the gene family members and (4) the ‘chimeric’ nature of B19.3 and B19.4, reflecting a cross-over or gene-conversion event in their common ancestor. We also show evidence for the presence of one or two additional expressed B19 genes in the barley genome. Based on our results, we present a model for the evolution of the family in barley, including the 20 amino acid motif. Comparisons of the relatedness between the barley family and all other known Group 1 Lea genes using maximum parsimony (PAUP) analysis provide evidence for the time of divergence between the barley genes containing the internal motif as a single copy and as a repeat. The PAUP analyses also provide evidence for independent duplications of Group 1 genes containing the internal motif as a repeat in both monocots and dicots.
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    Evolutionary origin of expandable G-C-rich triplet repeat DNA sequences (1995)
    Springer
    Biochemical genetics 33 (1995), S. 173-181 
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    ISSN: 1573-4927
    Keywords: fragile-X DNA systems ; expandable triplet repeats ; dynamic mutations ; conserved genetic domains ; evolution ; heritable disease mechanism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A model explaining properties exhibited by fragile-X DNA systems arises from observations that time-dependent base substitutions are expressed at G-C sites but not at A–T sites (Biochem. Genet.32:383, 1994). [CGG]n sequences are classified as most sensitive to evolutionary base substitution processes involving time-dependent populating of G-C sites with enol-imine states having enhanced stability. Increased density of these states in oocyte DNA would introduce a ground-state collapse double-helix of reduced energy that would inhibit strand separation by the replicase. Evolutionarily altered G′ in CG′G triplets allows CG′G to be transcribed as CTG, an initiation codon. And this will cause reinitiation of DNA synthesis, thereby adding additional CGG units to the collapsed double helix. This situation would not occur in slower-evolving male haploid DNA that replicates frequently.
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    Allozymic characterization and evolutionary relationships in the BrazilianAkodon cursor species group (Rodentia-Cricetidae) (1995)
    Springer
    Biochemical genetics 33 (1995), S. 283-295 
    Details
    ISSN: 1573-4927
    Keywords: Akodon ; Cricetidae rodents ; genetic diversity ; biochemical polymorphism ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The present study involved an electrophoretic survey of 22 protein loci in 269 individuals belonging to three species of the genusAkodon, A. aff.cursor (2n=16),A. cursor (2n=14/15), andA. montensis (2n=24/25/26), collected in Eastern Brazil. The joint results of gene diversity, genetic distances, phenetic analyses, and phylogenetic trees suggested thatA. aff.cursor has recently separated fromA. cursor and that the three species have experienced a recent chromosomal divergence followed by low allozyme differentiation. These data are in agreement with their classification as sibling species.
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    The primary structure ofHarpalus rufipes 5S ribosomal RNA (1995)
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    Molecular biology reports 21 (1995), S. 165-167 
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    ISSN: 1573-4978
    Keywords: 5S ribosomal RNA ; Harpalus rufipes ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleotide sequence of 5S ribosomal RNA from the beetleHarpalus rufipes was determined and compared with primary structures of other insect 5S rRNAs. Sequence differences between two beetle 5S rRNAs may represent phylogenetic markers specific for two groups of Coleoptera — Adephaga and Polyphaga. Analysis of all insect sequences using parsimony allowed us to infer a phylogenetic tree of insects, which is consistent with morphological and paleobiological data.
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    The two-hybrid: anin vivo protein-protein interaction assay (1995)
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    Molecular biology reports 21 (1995), S. 119-127 
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    ISSN: 1573-4978
    Keywords: β-galactosidase ; fusion protein ; protein-protein interaction ; Saccharomyces cerevisiae ; transcriptional activation ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    The myrosinase gene family in Arabidopsis thaliana: gene organization, expression and evolution (1995)
    Springer
    Plant molecular biology 27 (1995), S. 911-922 
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    ISSN: 1573-5028
    Keywords: Arabidopsis ; evolution ; expression ; genomic clone ; in situ hybridization ; myrosinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Myrosinase (thioglucoside glucohydrolase, EC 3.2.3.1.) is in Brassicaceae species such as Brassica napus and Sinapis alba encoded by two differentially expressed gene families, MA and MB, consisting of about 4 and 10 genes, respectively. Southern blot analysis showed that Arabidopsis thaliana contains three myrosinase genes. These genes were isolated from a genomic library and two of them, TGG1 and TGG2, were sequenced. They were found to be located in an inverted mode with their 3′ ends 4.4 kb apart. Their organization was highly conserved with 12 exons and 11 short introns. Comparison of nucleotide sequences of TGG1 and TGG2 exons revealed an overall 75% similarity. In contrast, the overall nucleotide sequence similarity in introns was only 42%. In intron 1 the unusual 5′ splice border GC was used. Phylogenetic analyses using both distance matrix and parsimony programs suggested that the Arabidopsis genes could not be grouped with either MA or MB genes. Consequently, these two gene families arose only after Arabidopsis had diverged from the other Brassicaceae species. In situ hybridization experiments showed that TGG1 and TGG2 expressing cells are present in leaf, sepal, petal, and gynoecium. In developing seeds, a few cells reacting with the TGG1 probe, but not with the TGG2 probe, were found indicating a partly different expression of these genes.
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    Two copies of a DNA element, ‘Wendy’, in the chloroplast chromosome of Chlamydomonas reinhardtii between rearranged gene clusters (1995)
    Springer
    Plant molecular biology 29 (1995), S. 63-80 
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    ISSN: 1573-5028
    Keywords: evolution ; Chlamydomonas reinhardtii ; chloroplast ; site-specific recombination ; transcription ; transposition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized two copies of a 2.4 kb DNA element that we call ‘Wendy’, in the chloroplast chromosome of Chlamydomonas reinhardtii. The two copies of Wendy reside in different single-copy regions at opposite positions in the chloroplast genome. Like many mobile DNA elements, both copies of Wendy are bordered by inverted repeats and contain several additional degenerate copies of these repeat sequences in direct or inverted orientation. In addition, four basepairs are repeated in direct orientation. Two major open reading frames (ORFs) are predicted from the DNA sequence of Wendy I. These ORFs are co-transcribed from a promoter inside the element. The deduced amino acid sequence of the larger of these ORFs shares some weak similarities with sequence motifs of transposases and integrases of other mobile elements. Wendy II appears to be altered relative to Wendy I by point mutations and small deletions and insertions which destroy the ORFs. The leader sequence of the Wendy transcript is nearly identical with the leader sequence of the rbcL transcript of C. reinhardtii, but not of C. moewusii (where the complete Wendy was also undetectable). Furthermore, both copies of Wendy are bracketed by gene clusters that are separated in C. reinhardtii but are contiguous in C. moewusii where they exist in an inverted orientation compared with C. reinhardtii. Wendy was not found in any of the completely sequenced chloroplast genomes of rice, tobacco, pine, Euglena or Marchantia, nor in any other GenBank entry. Our results suggest that Wendy has invaded C. reinhardtii after divergence from other species. Subsequent Wendy-dependent illegitimate homologous or site-specific recombination events or both may have contributed to scrambling of the C. reinhardtii chloroplast genome relative to genomes of other species.
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    URL: http://dx.doi.org/10.1007/BF00019119
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    A temperature-sensitive lambda cl repressor functions on a modified operator in yeast cells by masking the TATA element (1995)
    Springer
    Molecular genetics and genomics 248 (1995), S. 499-505 
    Details
    ISSN: 1617-4623
    Keywords: α-Amylase ; Glyceraldehyde-3-phosphate-dehydrogenase promoter ; Phage lambda ; Repression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe the construction and analysis of derivatives of the yeastTDH3 promoter in which the TATA box element has been replaced by a portion of the phage lambda operator containing a consensus TATA site flanked by binding sites for the cl repressor. Transcription of a reporter gene under the control of such a promoter is reduced in cells that express the cl repressor protein. Deletion of the native TATA element of theTDH3 promoter reduces transcription to the same extent. The cl repressor may act by “masking” the TATA element located between the repressor binding sites. Furthermore, the use of a temperature-sensitive cl repressor allowed temperature-dependent transcription of the reporter gene.
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    URL: http://dx.doi.org/10.1007/BF02191651
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    Isolation of temperature-sensitive mutants for mRNA capping enzyme in Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 147-154 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; CEG1 ; Temperature-sensitive mutants ; mRNA capping enzyme ; Guanylyltransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The guanylyltransferase activity of mRNA capping enzyme catalyzes the transfer of GMP from GTP to the 5′ terminus of mRNA. In Saccharomyces cerevisiae, the activity is carried on the α subunit of capping enzyme, the product of the CEG1 gene. We have isolated 10 recessive, temperature-sensitive mutations of CEG1; nine (cegl-1 to cegl-9) were isolated on a single-copy plasmid and the remaining one (cegl-10) on a multicopy plasmid. The presence of cegl-10 in multiple copies is essential for the viability of cells carrying the mutation, and a shift to the restrictive temperature resulted in rapid growth arrest of cegl-10 cells, while growth rates of other mutants decreased gradually upon temperature upshift. Intragenic complementation was not observed for pairwise combinations of the mutations. Although the majority of the mutations occurred at the amino acid residues conserved between Cegl and the Schizosaccharomyces pombe homologue, none were located in the regions that are also conserved among viral capping enzymes and polynucleotide ligases. Guanylyltransferase activity of the mutant proteins as measured by covalent Ceg1-GMP complex formation was heat-labile. The availability of these mutants should facilitate studies of the structure-function relationships of capping enzyme, as well as the roles and regulation of mRNA capping.
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    URL: http://dx.doi.org/10.1007/BF00290360
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    Enhanced stability in vivo of a thermodynamically stable mutant form of yeast iso-1-cytochrome c (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 155-161 
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    ISSN: 1617-4623
    Keywords: Cytochrome c ; Protein stability ; Protein degradation ; Mitochondria ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previous work has established that the N57I amino acid replacement in iso-1-cytochrome c from the yeast Saccharomyces cerevisiae causes an unprecedented increase in thermodynamic stability of the protein in vitro, whereas the N57G replacement diminishes stability. Spectrophotometric measurements of intact cells revealed that the N57I iso-l-cytochrome c is present at higher than normal levels in vivo. Although iso-1-cytochrome c turnover is negligible during aerobic growth, transfer of fully derepressed, aerobically grown cells to anaerobic growth conditions leads to reduction in the levels of all of the cytochromes. Pulsechase experiments carried out under these anaerobic conditions demonstrated that the N57I iso-l-cytochrome c has a longer half-life than the normal protein. This is the first report of enhanced stability in vivo of a mutant form of a protein that has an enhanced thermodynamic stability in vitro. Although the N57I protein concentration is higher than the normal level, reduced growth in lactate medium indicated that the specific activity of this iso-l-cytochrome c in vivo is diminished relative to wild-type. On the other hand, the level of the thermodynamically labile N57G iso-1-cytochrome c was below normal. The in vivo levels of the N57I and N57G iso-l-cytochrome c suggest that proteins in the mitochondrial intermembrane space can be subjected to degradation, and that this degradation may play a role in controlling their normal levels.
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    A C-terminal region of the Saccharomyces cerevisiae transcription factor ADR1 plays an important role in the regulation of peroxisome proliferation by fatty acids (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 289-296 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Peroxisomes ; Catalase A ; ADR1 ; Peroxisome proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Saccharomyces cerevisiae transcriptional activator ADR1, which controls ADH2 gene expression, was shown to be involved in the regulation of peroxisome proliferation. To study the mode of action of ADR1, we compared strains carrying the adr1-1 mutation, high or low copy numbers of the ADR1 gene, the constitutive allele ADR1-5 c, and 3′-deletions of ADR1. High ADR1 gene dosage increased the transcription of genes encoding peroxisomal proteins as compared to one copy of the ADR1 gene. Furthermore, overexpression of ADR1 under ethanol growth conditions induced the proliferation of peroxisomal structures. The organelles were observed to be localized in clusters, a typical feature of peroxisomes induced by oleic acid. In contrast, the ADR1-5 c allele, which induces ADH2 expression to a level comparable to that of high ADR1 gene dosage was found to have only a small effect. An analysis of functional domains of the ADR1 protein revealed that the N-terminal 220 amino acids of ADR1 were sufficient for wild-type levels of transcription of the FOX2, FOX3, and PAS1 genes, but the entire ADR1 protein was required for complete induction of the CTA1 gene and for growth oleic acid medium. Our data suggest that a functional domain of the ADR1 protein localized between residues 643 and 1323 is required for the induction of peroxisomal structures and for the utilization of oleic acid.
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    Cloning and characterization of seven cDNAs for hyperosmolarity-responsive (HOR) genes of Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 127-138 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Hyperosmotic stress ; Signal transduction pathway
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Yeast cells can respond and adapt to osmotic stress. In our attempt to clarify the molecular mechanisms of cellular responses to osmotic stress, we cloned seven cDNAs for hyperosmolarity-responsive (HOR) genes from Saccharomyces cerevisiae by a differential screening method. Structural analysis of the clones revealed that those designated HOR1, HORS, HOR4, HOR5 and HOR6 encoded glycerol-3-phosphate dehydrogenase (Gpd1p), glucokinase (Glklp), hexose transporter (Hxtlp), heat-shock protein 12 (Hsp12p) and Na+, K+, Li+-ATPase (Enalp), respectively. HOR2 and HOR7 corresponded to novel genes. Gpdlp is a key enzyme in the synthesis of glycerol, which is a major osmoprotectant in S. cerevisiae. Cloning of HOR1/GPD1 as a HOR gene indicates that the accumulation of glycerol in yeast cells under hyperosmotic stress is, at least in part, caused by an increase in the level of GPDH protein. We performed a series of Northern blot analyses using HOR cDNAs as probes and RNAs prepared from cells grown under various conditions and from various mutant cells. The results suggested that all the HOR genes are regulated by common signal transduction pathways. However, the fact that they exhibited certain distinct responses indicated that they might also be regulated by specific pathways in addition to the common pathways. Ca2+ seemed to be involved in the signaling systems. In addition, Hog1p, one of the MAP kinases in yeast, appeared to be involved in the regulation of expression of HOR genes, although its function seemed to be insufficient for the overall regulation of expression of these genes.
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    Rapid and extensive vacuolation of the budding yeastsSaccharomyces cerevisiae andCandida albicans by amphotericin B (1995)
    Springer
    Mycoscience 36 (1995), S. 147-150 
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    ISSN: 1618-2545
    Keywords: amphotericin B ; budding yeasts ; Candida albicans ; Saccharomyces cerevisiae ; vacuolation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of amphotericin B (AMPH) on vacuolation in the budding yeastsSaccharomyces cerevisiae andCandida albicans was studied. The minimum inhibitory concentration of AMPH for growth ofS. cerevisiae andC. albicans was 1 µg/ml. In untreated control cultures, mature cells had large central vacuoles in the exponential phase, which hampered the detection of vacuolation effect. Small buds in untreated exponential phase cells, however, only rarely showed vacuoles under the light microscope. Treatment with 0.2 µg/ml of AMPH for 20–30 min induced extensive vacuolation not only in mothers but also buds ofS. cerevisiae. Extensive vacuolation lasted 4 h or more, and growth rate of the cells was much reduced for 8 h or more. Vacuolation itself was not fatal: on removal of the drug most cells gradually recovered from vacuolation and eventually multiplied. A similar effect of AMPH was also observed inC. albicans but at a higher concentration (0.5 µg/ml).
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    Isolation and characterization of extragenic mutations affecting the expression of the uroporphyrinogen decarboxylase gene (HEM12) in Sacharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 247 (1995), S. 471-481 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Uroporphyrinogen decarboxylase ; HEM12 transcription ; Porphyria cutanea tarda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Uroporphyrinogen decarboxylase (Uro-d; EC 4.1.1.37), the fifth enzyme in the heme biosynthetic pathway, which catalyzes the sequential decarboxylation of uroporphyrinogen to coproporphyrinogen, is encoded by the HEM12 gene in Saccharomyces cerevisiae. The HEM12 gene is transcribed into a major short mRNA and a minor longer one, approximately 1.35 and 1.55 kb, respectively, in size, and that differ in the 5′ untranslated region. “Uroporphyric” mutants, which have no mutations in the HEM12 gene but accumulate uroporphyrinogen, a phenotype chracteristic of partial Uro-d deficiency, were investigated. Genetic analysis showed that the mutant phenotype depends on the combined action of two unlinked mutations, udt1 and either ipa1, ipa2, or ipa3. ipa1 is tightly linked to HEM12 The mutation udt1 apparently acts specifically on the HEM12 gene, and causes a six to tenfold decrease in the levels of the short HEM12 mRNA, in the β-galactosidase activity of a HEM12-lacZ fusion, in immunodetectable protein and enzyme activity. But heme synthesis is normal and porphyrin accumulation was modest. The mutations ipa1, ipa2, and ipa3 had no phenotype on their own, but they caused an increase in porphyrin accumulation in a udt1 background. This multiplicity of genetic factors leading to uroporphyric yeast cells closely resembles the situation in human porphyria cutanea tarda.
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    On the mechanism of UV and γ-ray-induced intrachromosomal recombination in yeast cells synchronized in different stages of the cell cycle (1995)
    Springer
    Molecular genetics and genomics 248 (1995), S. 301-310 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Intrachromosomal recombination ; Cell cycle ; Radiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genetic system selecting for deletion events (DEL recombination) due to intrachromosomal recombination has previously been constructed in the yeastSaccharomyces cerevisiae. Intrachromosomal recombination is inducible by chemical and physical carcinogens. We wanted to understand better the mechanism of induced DEL recombination and to attempt to determine in which phase of the cell cycle DEL recombination is inducible. Yeast cells were arrested at specific phases of the cell cycle, irradiated with UV or γ-rays, and assayed for DEL recombination and interchromosomal recombination. In addition, the contribution of intrachromatid crossing-over to the number of radiation induced DEL recombination events was directly investigated at different phases of the cell cycle. UV irradiation induced DEL recombination preferentially in S phase, while γ-rays induced DEL recombination in every phase of the cell cycle including G1. UV and γ-radiation induced intrachromatid crossing over preferentially in G1, but it accounted at the most for only 14% of the induced DEL recombination events. The possibility is discussed that single-strand annealing or one-sided invasion events, which can occur in G1 and may be induced by a double-strand break intermediate, may be responsible for a large proportion of the induced DEL recombination events.
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    URL: http://dx.doi.org/10.1007/BF02191597
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    The pollinium ofLoroglossum hircinum (Orchidaceae) between pollination and pollen tube emission (1995)
    Springer
    Plant systematics and evolution 196 (1995), S. 141-151 
    Details
    ISSN: 1615-6110
    Keywords: Orchidaceae ; Loroglossum hircinum ; Compound pollen ; pollinium ; pollen tubes ; generative cell ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The structure of the massulae composing the pollinium ofLoroglossum hircinum was studied before pollination and 12 and 24 hours afterwards. The grains are grouped in tetrads closely packed in massulae. The exine is only present on the outside of the massulae. The intine consists of two layers: a compact layer surrounding the pollen grain and a looser layer surrounding the pollen grain and a looser layer surrounding the tetrad. Twelve hours after pollination, pollen volume and the space between the tetrads increase due to vacuolization. Twenty-four hours after pollination, pollen volume and tetrad spacing are higher due to vacuolization and some grains have emitted pollen tubes. Pollen growth due to vacuole formation, and the absence of common walls between adjacent tetrads lead to crumbling of the massulae. The mature pollen grain does not have apertures: the site of pollen tube emission is determined after pollination. The first grains to germinate are those in the centre of the massula. The vegetative cell nucleus is the first to enter the pollen tube; the generative cell elongates and undergoes the second haploid mitosis shortly after entering the pollen tube.
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    Genetic relationships between peanut and wild species ofArachis sect.Arachis (Fabaceae): Evidence from RAPDs (1995)
    Springer
    Plant systematics and evolution 198 (1995), S. 167-178 
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    ISSN: 1615-6110
    Keywords: Fabaceae ; Arachis ; Arachis hypogaea ; RAPD ; systematics ; evolution ; germplasm resources
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Twenty-six accessions of wildArachis species and domesticated peanuts,A. hypogaea, introduced from South America were analyzed for random amplified polymorphic DNA (RAPD). The objective of the study was to investigate inter- and intraspecific variation and affinities among species of sect.Arachis which have been proposed as possible progenitors for the domesticated peanut. Ten primers resolved 132 DNA bands which were useful for separating species and accessions. The most variation was observed among accessions ofA. cardenasii andA. glandulifera whereas the least amount of variation was observed inA. hypogaea andA. monticola. The two tetraploid species could not be separated by using RAPDs.Arachis duranensis was most closely related to the domesticated peanut and is believed to be the donor of the A genome. The data indicated thatA. batizocoi, a species previously hypothesized to contribute the B genome toA. hypogaea, was not involved in its evolution. The investigation showed that RAPDs can be used to analyze both inter- and intraspecific variation in peanut species. Southern hybridization of RAPD probes to blots containing RAPD of theArachis species provided information on genomic relationships and revealed the repetitive nature of the amplified DNA.
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    How is a species kept together? (1995)
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    Biology and philosophy 10 (1995), S. 181-196 
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    ISSN: 1572-8404
    Keywords: Biological species concept ; gene flow ; gene circulation ; Ernst Mayr ; stalemates ; typology ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Over the decades, there has been substantial empirical evidence showing that the unity of species cannot be maintained by gene flow. The biological species concept is inconclusive on this point. The suggestion is made that the unity of species is maintained rather by selection constantly spreading new alleles throughout the species, or bygene circulation. There is a lack in conceptual distinction between gene flow and gene circulation which lies at the heart of the problem. The concept of gene circulation also sheds some new light on the problem of typology and on such a broad concept as evolution. A new species definition is proposed.
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    A (not-so-radical) solution to the species problem (1995)
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    Biology and philosophy 10 (1995), S. 339-356 
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    ISSN: 1572-8404
    Keywords: Species ; lineage ; individual ; class ; evolution ; organism ; population
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract What are species? One popular answer is that species are individuals. Here I develop another approach to thinking about species, an approach based on the notion of a lineage. A lineage is a sequence of reproducing entities, individuated in terms of its components. I argue that one can conceive of species as groups of lineages, either organism lineages or population lineages. Conceiving of species as groups of lineages resolves the problems that the individual conception of species is supposed to resolve. It has added the virtue of focusing attention on the characteristic of species that is most relevant to understanding their role in evolutionary processes, namely, the lineage structure of species.
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    Viability explanation (1995)
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    Biology and philosophy 10 (1995), S. 435-457 
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    ISSN: 1572-8404
    Keywords: Functional explanation ; morphology ; ethology ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract This article deals with a type of functional explanation, viability explanation, that has been overlooked in recent philosophy of science. Viability explanations relate traits of organisms and their environments in terms of what an individual needs to survive and reproduce. I show that viability explanations are neither causal nor historical and that, therefore, they should be accounted for as a distinct type of explanation.
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    Are the different hypotheses on the emergence of life as different as they seem? (1995)
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    Biology and philosophy 10 (1995), S. 389-417 
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    ISSN: 1572-8404
    Keywords: Catalysis ; chance ; determinism ; emergence of life ; evolution ; non-equilibrium thermodynamics ; panspermia ; protometabolism ; reduction ; RNA world ; self-organization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract This paper calls attention to a philosophical presupposition, coined here “the continuity thesis” which underlies and unites the different, often conflicting, hypotheses in the origin of life field. This presupposition, a necessary condition for any scientific investigation of the origin of life problem, has two components. First, it contends that there is no unbridgeable gap between inorganic matter and life. Second, it regards the emergence of life as a highly probable process. Examining several current origin-of-life theories. I indicate the implicit or explicit role played by the “continuity thesis” in each of them. In addition, I identify the rivals of the “thesis” within the scientific community — “the almost miracle camp.” Though adopting the anti-vitalistic aspect of the “continuity thesis”, this camp regards the emergence of life as involving highly improbable events. Since it seems that the chemistry of the prebiotic stages and of molecular self-organization processes rules out the possibility that life is the result of a “happy accident,” I claim that the “almost miracle” view implies in fact, a creationist position.
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    Glycan modification of a thermostable recombinant (1-3,1-4)-β-glucanase secreted fromSaccharomyces cerevisiae is determined by strain and culture conditions (1995)
    Springer
    Glycoconjugate journal 12 (1995), S. 380-390 
    Details
    ISSN: 1573-4986
    Keywords: glycosylation ; Saccharomyces cerevisiae ; heterologous ; glucanase ; thermostability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract High level biosynthesis and secretion of the thermostable hybrid (1-3,1-4)-β-glucanase H(A16-M) has been achieved inSaccharomyces cerevisiae by means of the yeast vacuolar endoprotease B promoter (PRB1p) and theBacillus macerans (1-3,1-4)-β-glucanase signal peptide. The N-glycans present on the yeast-secreted H(A16-M), denoted H(A16-M)-Y, were released by endoglycosidase H, and identified by proton NMR spectroscopy to be a homologous series of Man8-13GlcNAc2, although only traces of Man9GlcNAc2 were found. Therefore, processing of N-glycans on H(A16-M)-Y is similar to that on homologous proteins. Most of the N-glycans (88%) were neutral while the remainder were charged due to phosphorylation. Site-directed mutagenesis of Asn to Gln in two of the N-glycosylation sequons, and subsequent analysis of the N-glycans on the yeast-secreted proteins together with analysis of the N-glycans from the individual sites of H(A16-M)-Y suggest the presence of steric hindrance to glycan modification by the glycans themselves. H(A16-M)-Y produced under control of either the yeast protease B or the yeast 3′-phosphoglycerate kinase promoter, each in two differentSaccharomyces strains revealed a dependence of N-glycan profile on both strain and culture conditions. The extent of O-glycosylation was found to be nine mannose units per H(A16-M)-Y molecule. An attempt to identify the linkage-sites for the O-glycans by amino acid sequencing failed, suggesting non-stoichiometric or heterogeneous O-glycosylation. The possible modes in which N-glycans might contribute to resistance of H(A16-M)-Y to irreversible thermal denaturation are discussed with respect to structural information available for H(A16-M)-Y.
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    Genetic diversity, distributional barriers and rafting continents — more thoughts on the evolution of mangroves (1995)
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    Hydrobiologia 295 (1995), S. 167-181 
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    ISSN: 1573-5117
    Keywords: mangrove ; Avicennia ; evolution ; fossils ; pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Without continental drift, the diversity and distribution of many species, including mangrove plants, would be very different today. First, there would be fewer pantropic genera and many more endemics. Second, their characteristics would not be as common and widespread as some are today. Continental drift has brought about the massive mixing and dispersal of genes in geologically recent times, greatly enhancing the evolutionary process; particularly for flowering plants — the angiosperms, which evolved during the period. Mangrove plants are comprised of approximately 70 species from 20 quite different angiosperm families. Most taxa are characterized by special physiological abilities and structural forms, enabling them to live in both seasonally fluctuating saline conditions, and water-saturated soils. Their occurrence is mostly tropical, perhaps because of harsh physiological conditions of intertidal habitats; but distributions of specific taxa do not fully concur with the idea of a completely tropical evolution, at least for some important species. At least one genus of mangrove tree, Avicennia, occurs around the world, chiefly in tropical estuarine habitats, although they also range into temperate latitudes, especially in the south. Around the world, there are no more than ten species of Avicennia recognised today, but their diagnostic determinants were inadequate prior to recent studies using both numerical analyses of morphological parameters and isozymes. Such analyses significantly reduced the number of apparent species, notably around Australia, and provided a basis for the revision of distributional records throughout the Indo West Pacific region. One species, A. marina, was found to be widespread and morphologically variable with genes divided into characteristic groupings of at least three geographic areas in the region. Based on these findings, there are several novel inferences to be made regarding the evolution of this genus. A western Gondwanan origin is proposed, with subsequent radiation of several taxa facilitated via the tectonic dispersal of southern continental fragments.
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    The evolution of groundwater Cladocera (1995)
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    Hydrobiologia 307 (1995), S. 69-74 
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    ISSN: 1573-5117
    Keywords: groundwater ; evolution ; Cladocera ; Alona ; conserved species
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cladocera occur in various types of groundwater, but are most common in the underflow of rivers. Numerous surface water species occasionally venture into groundwater; some chydorids are more common in groundwaters than in surface waters; at least three groups within Alona, finally, have evolved exclusive groundwater species. The latter show few obvious adaptations to the subterranean habitat, except loss of an eye or total blindness. Some, however, have conserved an array of primitive characters (e.g. on the end-claw of the postabdomen, and the setation of the valve rims) which suggest that the physical protection and relative constancy of the hyporheic has permitted the survival of some ancient taxa.
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    An isolated population of fourhorn sculpins (Myoxocephalus quadricornis, family Cottidae) in a hypersaline high arctic Canadian lake (1995)
    Springer
    Hydrobiologia 312 (1995), S. 27-35 
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    ISSN: 1573-5117
    Keywords: mining impacts ; sculpins ; cephalic spines ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Freshwater sculpins probably evolved from marine ancestors which entered bodies of water such as proglacial lakes or lakes which were gradually isolated from the sea by isostatic rebound. Sculpins in fresh water lakes (Myoxocephalus thompsoni [Girard]) lack cephalic horns and live well below a depth of 10 m. Those in the sea (Myoxocephalus quadricornis [Linnaeus]) typically live above 10 m and possess a well developed set of four cephalic horns. The sculpins in Garrow Lake, North West Territories, are intermediate between the marine and fresh water forms with respect to their depth distributions and their cephalic horns (spines). As a consequence, Garrow Lake, which separated from the sea some 3000 years ago, serves as an excellent ‘laboratory’ for studying evolutionary changes in this sculpin. The age of the lake was based on carbon-14 dates of the fossil pelecypods from raised beaches around the lake and from observations of rates of isostatic rebound in the area as reported by Dickman & Ouellet 1983 and Pagé et al. 1984. During the last 3000 years, the surface waters of Garrow Lake have freshened and its sculpins have apparently adapted to this top down freshening by occupying a depth where the salinity of the lake approaches that of sea water. As a result, the sculpin population in Garrow Lake lives deeper than the sculpin population in the nearby Garrow Bay. Thus, the deeper dwelling Garrow Lake sculpins appear to be less vulnerable to avian predation than their shallow water dwelling marine ancestors. It is hypothesized that reduced avian predation of the Garrow Lake sculpin population is associated with the observed reduction in their cephalic horns which impart a certain degree of disruptive colouration and disruptive pattern outline allowing the shallow dwelling marine species to blend in with its background in a manner which appears to make it less visible to avian predators. It is unfortunate that the three thousand year old Garrow Lake sculpin population is now endangered by mine tailings entering the lake from the nearby Cominco Ltd. mine. The entire food chain of the lake appears to have been severely impacted by lead and zinc mine tailings entering Garrow Lake at a rate of 100 metric tonnes per hour.
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    URL: http://dx.doi.org/10.1007/BF00018884
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    Phylogeny and adaptive radiation within the Anomopoda: a preliminary exploration (1995)
    Springer
    Hydrobiologia 307 (1995), S. 57-68 
    Details
    ISSN: 1573-5117
    Keywords: Anomopoda ; evolution ; phylogeny ; adaptive radiation ; morphology ; ecology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The distinctness of the Anomopoda and the polyphyletic nature of the so-called Cladocera are emphasized. An attempt is made to reconstruct the ancestral anomopod, which probably lived in Palaeozoic times. This task is facilitated by the availability of detailed information on extant forms, which includes functional as well as purely morphological considerations and enables us to understand the means whereby complex mechanisms were transformed during evolution. Comparative studies on the ecology and habits of extant forms also throw light on the probable way of life of the ancestral anomopod. Adaptive radiation within the Anomopoda is briefly surveyed and an outline of the suggested phylogeny of the order is indicated.
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    URL: http://dx.doi.org/10.1007/BF00031997
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    The late Cenozoic ampullariidae (mollusca, gastropoda) of the Albertine Rift Valley (Uganda-Zaire) (1995)
    Springer
    Hydrobiologia 316 (1995), S. 1-32 
    Details
    ISSN: 1573-5117
    Keywords: Africa ; autapomorphic characters ; convergence ; evolution ; freshwaters snails ; Lanistes ; Mollusca ; palaeolimnology ; palaeontology ; Pila ; predator/prey coevolution ; riftlakes ; taxonomy ; punctuated equilibrium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Albertine Rift Valley (Uganda-Zaire) contains vast sedimentary sequences of late Cenozoic age. They were deposited in an extensive riftlake, Lake Obweruka, which existed from c. 8 Ma to 2.5 Ma and was comparable in size and depth to the present L. Tanganyika. Many freshwater molluscs that occur in these lacustrine deposits are characterised by their aberrant shell morphology, their extreme ornamentation and general form, making them resemble marine species. This convergence, rare in freshwater molluscs, is called thalassoidism and extreme ornamentation in marine as well as in freshwater molluscs is considered to be the result of a gradual process of prey/predator coevolution. In the present paper the Albertine representants of the ampullariid genera Lanistes and Pila, most of which are new to science, are taxonomically described and their phylogenetic relation, based upon apomorphic characters, is given. In addition the evolutionary history of these freshwater snails in the basin has been reconstructed. In the pre-riftlake environment 3 species of Lanistes occurred, with no special shell adaptations against predation. After the formation of a riftlake, 2 of these, colonising the new lacustrine ecospace, changed morphologically and radiated. The 3 derived lines show minor adaptations against predation. After the extinction of the dominant Lanistes species group around 6 Ma, the sole surviving lacustrine Lanistes suddenly radiates, the ancestral line persisting next to the 3 new daughter lines. This second morphological shift is spectacular as it produces shells with distinct thalassoid features. All the Lanistes species of L. Obweruka became extinct during a cataclysmic event around 4.5 Ma. Populations of the genus Pila colonised lacustrine habitats after this event, the derived form also showing striking thalassoid characters. There is no doubt that the intense morphological change occurred during a brief period, geologically speaking. The degree of morphological change in molluscs appears hence not to be linked with time. After the sudden radiation all lineages remain morphologically stable until they became extinct c. 1 Ma later. This pattern corresponds to the punctuated equilibrium model. Other groups (viviparids, thiarids) show more gradual changes.
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    Effects of L-alanine and L-alanine peptides on the chemotaxis of tetrahymena: Evolutionary conclusions (1995)
    Springer
    Bioscience reports 15 (1995), S. 185-190 
    Details
    ISSN: 1573-4935
    Keywords: L-alanine ; evolution ; chemosensory response ; peptides ; imprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract L-alanine and its peptides (L-Ala-2–6) do not attract or repulse Tetrahymena in a 10−8M concentration. In 10−10M concentration there is a consistent repellent effect. Twenty four hours after L-alanine or L-alanine-peptides' pretreatment (imprinting) the progeny generation of the cells react differently to the same materials. L-Alanine, L-alanine penta- and hexapeptide in both concentrations are chemoattractant, while L-alanine tetrapeptide is repellent. L-Alanine dipeptide is inert in 10−10M and repellent at 10−8M concentrations, while L-alanine tripeptide is strongly repellent at 10−10M and attractant at 10−8M concentrations. This means, that the first encounter (imprinting) with an exogeneous amino acid or peptide is decisive to the later reaction of the protozoan cell. The chain length is important in the imprinting, however the reaction is not consistent. The experiments call the attention to the significance of imprinting in the receptor and hormone evolution.
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    Demography of source—sink populations and the evolution of ecological niches (1995)
    Springer
    Evolutionary ecology 9 (1995), S. 38-44 
    Details
    ISSN: 1573-8477
    Keywords: demography ; dispersal ; ecological niche ; evolution ; heterogeneous environments ; natural selection ; source—sink populations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The demography of populations living in variable environments is an important factor molding the evolution of ecological niches, for it determines the relative strength of selection pressures on adaptations to different habitats. Here I consider a coarse-grained environment consisting of two habitat types and investigate how the selection pressure on reproductive success in different habitats depends on their quality and frequency and the dispersal pattern. The results suggest that selection on adaptations to optimal habitats will usually be stronger than on adaptations to poor habitats and the ecological niche will thus tend to be an evolutionarily conservative character. It is because under the habitat choice or limited dispersal that seem to prevail in natural populations, more individuals encounter the better habitat than would be expected solely on the basis of its relative area. This bias results in reduced selection pressure on reproductive success in the poorer habitat. With habitat choice or limited dispersal, selection pressure on reproductive success in the poorer habitat may exceed that on reproductive success in the better habitat only if the poorer habitat is much more frequent in the environment than the better habitat and the difference in their quality is not large.
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    URL: http://dx.doi.org/10.1007/BF01237695
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    Actin cytoskeleton and budding pattern are altered in the yeast rvs161 mutant: the Rvs161 protein shares common domains with the brain protein amphiphysin (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 485-495 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Actin cytoskeleton ; Budding pattern ; Amphiphysin ; Myosin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The actin cytoskeleton cells is altered in rvs161 mutant yeast, with the defect becoming more pronounced under unfavorable growth conditions, as described for the rvs167 mutant. The cytoskeletal alteration has no apparent effect on invertase secretion and polarized growth. Mutations in RTVS161, just as in RI/S167, lead to a random budding pattern in a/α diploid cells. This behavior is not observed in a/a diploid cells homozygous for the rvs161-1 or rvs167-1 mutations. In addition, sequence comparisons revealed that amphiphysin, a protein first found in synaptic vesicles of chicken and shown to be the autoantigen of Stiff Man syndrome, presents similarity with both Rvs proteins. Furthermore, limited similarities with myosin heavy chain and tropomyosin alpha chain from higher eukaryotic cells allow for the definition of a possible consensus sequence. The finding of related sequences suggests the existence of a function for these proteins that is conserved among eukaryotic organisms.
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    Evidence that two zinc fingers in the methionine aminopeptidase from Saccharomyces cerevisiae are important for normal growth (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 247-253 
    Details
    ISSN: 1617-4623
    Keywords: Methionine aminopeptidase (MAP) ; Metallopeptidase ; Zinc finger ; Mutagenesis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Limited proteolysis of intact yeast methionine aminopeptidase (MAP1) with trypsin releases a 34 kDa fragment whose NH2-terminal sequence begins at Asp70, immediately following Lys69. These results suggest that yeast MAP may have a two-domain structure consisting of an NH2-terminal zinc finger domain and a C-terminal catalytic domain. To test this, a mutant MAP lacking residues 2–69 was generated, overexpressed, purified and analyzed. Metal ion analyses indicate that 1 mol of wild-type yeast MAP contains 2 mol of zinc ions and at least 1 mol of cobalt ion, whereas 1 mol of the truncated MAP lacking the putative zinc fingers contains only a trace amount of zinc ions but still contains one mole of cobalt ion. These results suggest that the two zinc ions observed in the native yeast MAP are located at the Cys/His rich region and the cobalt ion is located in the catalytic domain. The k.at and Km values of the purified truncated MAP are similar to those of the wild-type MAP when measured with peptide substrates in vitro and it appears to be as active as the wild-type MAP in vivo. However, the truncated MAP is significantly less effective in rescuing the slow growth phenotype of map mutant than the wild-type MAP. These findings suggest that the zinc fingers are essential for normal MAP function in vivo, even though the in vitro enzyme assays indicate that they are not involved in catalysis. In addition, a series of single mutations were generated by changing the cysteines and the histidines in the zinc finger region to serines and arginines, respectively. Analyses of these point mutations provide further evidence that the cysteines and histidines are important for the growth promotion function of yeast MAP.
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    The CLS2 gene encodes a protein with multiple membrane-spanning domains that is important Ca2+ tolerance in yeast (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 269-281 
    Details
    ISSN: 1617-4623
    Keywords: Ca2+ homeostasis ; Saccharomyces cerevisiae ; CLS2 ; Endoplasmatic reticulum ; Membrane protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic screening of Saccharomyces cerevisiae mutants defective in Ca2+ homeostasis identified cls2, which exhibits a specific Ca2+-sensitive growth phenotype. We describe here the CLS2 gene and a multicopy suppressor (named BCL21, for bypass of CLS2) of the cls2 mutation. The CLS2 gene encodes a polypeptide of 410 amino acid residues, and its hydropathy profile indicates that the predicted Cls2 protein (Cls2p) contains ten putative membrane spanning regions. Immunofluorescent staining of the yeast cells expressing epitopetagged Cls2p suggests that Cls2p is localized to endoplasmatic reticulum (ER) membrane. A cls2 disruption strain is viable, but shows a Ca2+-sensitive phenotype like the original cls2 mutants. BCL21 suppresses the cls2 disruption mutation, indicating that the multicopy suppression does not require the Cls2p. Suppression of cls2 was observed even after introduction of a singlecopy plasmid harboring BCL21. The BCL21 gene encodes a protein of 382 amino acid residues and is identical to the SUR1 gene. sur1 was originally isolated as a suppressor of rvs161, which has reduced viability in nutrient starvation conditions. Possible mechanisms of the multicopy suppression are discussed.
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    Identification and characterization of regulatory elements in the phosphoenolpyruvate carboxykinase gene PCK1 of Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 367-373 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Phosphoenolpyruvate carboxykinase ; Glucose repression ; Gluconeogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phosphoenolpyruvate carboxykinase is a key enzyme in gluconeogenesis. The expression of the PCK1 gene in Saccharomyces cerevisiae is strictly regulated and dependent on the carbon source provided. Two upstream activation sites (UAS1PCK1 and UAS2PCK1) and one upstream repression site (URSPCK1) were localized by detailed deletion analysis. The efficacy of these three promoter elements when separated from each other was confirmed by investigations using heterologous promoter test plasmids. Activation mediated by UAS1PCK1 or UAS2PCK1 did not occur in the presence of glucose, indicating that these elements are essential for glucose derepression. The repressing effect caused by URSPCK1 was much stronger in glucose-grown cells than in ethanol-grown cells.
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    Cis and trans-acting regulatory elements required for regulation of the CPS1 gene in Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 580-589 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Gene regulation ; Carboxypeptidase yscS ; Vacuolar peptidase ; Regulatory elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To clarify the transcriptional regulation by nutrient limitation of the gene encoding carboxypeptidase yscS in Saccharomyces cerevisiae (CPS1), we performed an analysis of its 5′ noncoding region. In deletion experiments a sequence located between positions −644 and −591 was found to be responsible for transcriptional repression of the CPS1 gene in yeast cells grown on rich nitrogen sources. Furthermore, a 162 bp fragment spanning positions −644 to −482 of the promoter of the CPS1 gene repressed gene expression when placed 3′ to the upstream activation sequence (UAS) of the heterologous gene CYC1. A fragment containing this putative upstream repression sequence (URS) was shown specifically to bind protein from a yeast extract as demonstrated by gel retardation experiments. Although a sequence mediating the control of gene expression by GCN4 was found within the URS element, the GCN4 gene product is not required for DNA-binding activity. In addition, at least three other upstream activation UASs responsible for the activation of CPS1 expression by glucose under nitrogen starvation conditions were found to be located between positions −673 and −644, −482 and −353, and −243 and −186, respectively. The putative mechanism of the nitrogen limitation-dependent regulation of CPS1 expression via these regulatory elements is discussed.
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    Dosage suppressors of the dominant G1 cyclin mutantCLN3-2: Identification of a yeast gene encoding a putative RNA/ssDNA binding protein (1995)
    Springer
    Molecular genetics and genomics 248 (1995), S. 712-718 
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    ISSN: 1617-4623
    Keywords: G1 cyclin ; RNA binding protein ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three G1 cyclins,CLN1,CLN2, andCLN3, have been identified in the budding yeastSaccharomyces cerevisiae. G1 cyclins are essential, albeit functionally redundant, rate-limiting activators of cell cycle initiation. We have isolated dosage-dependent suppressor genes (designatedHMD genes) of the mating defect caused byCLN3-2, a dominant mutation inCLN3,HMD2 andHMD3 are identical toSTE4 andSTE5, respectively,HMD1 is an essential gene that encodes a protein containing a putative RNA binding domain. Overproduction ofHMD1 results in a relatively specific reduction in the level of theCLN3 orCLN3-2 transcript. This reduction occurs subsequent to transcription initiation ofCLN3 since overexpression ofHMD1 did not affect expression of a heterologous transcript from theCLN3 promoter but did result in a reduction ofCLN3 transcript expressed from a heterologous promoter.HMD1 has at least one essential role independent of its effect onCLN3 sinceHMD1 remains essential for viability in the absence of a functionalCLN3 gene.
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    Overexpression of the RNR1 gene rescues Saccharomyces cerevisiae mutants in the mitochondrial DNA polymerase-encoding MIP1 gene (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 1-7 
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    ISSN: 1617-4623
    Keywords: Polymerase ; Ribonucleotide reductase ; Mitochondrial DNA ; Saccharomyces cerevisiae ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A multicopy suppressor gene which rescues the temperature-sensitive growth defect of Saccharomyces cerevisiae mutants in the mitochondrial DNA (mtDNA) polymerase-encoding MIP1 gene has been isolated and identified as the RNR1 gene. This gene, whose transcript is cell cycle-regulated and mainly expressed at the G1 to S phase transition, encodes the large subunit of ribonucleotide reductase. This enzyme catalyses a limiting step in the production of deoxynucleotides needed for DNA synthesis. The presence of a high copy number of the RNR1 gene also decreases the accumulation of rho− mutants observed in diploids that harbour a single copy of the MIP1 gene. In cell cycle-synchronised cells, the presence of a high copy number of RNR1 does not modify its cell cycle transcription regulation and increases its transcript level by a factor of 10 throughout the cell cycle. Our results show that an increased supply of dNTPs in mitochondria can stimulate the mtDNA polymerase activity and indicate that the dNTP concentration may be rate limiting for the replication of mtDNA.
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    Artificial antisense RNA regulation of YBR1012 (YBR136w ), an essential gene from Saccharomyces cerevisiae which is important for progression through G1/S (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 51-57 
    Details
    ISSN: 1617-4623
    Keywords: Antisense RNA ; YBR1012/YBR136w/MEC1/ESR1 ; Saccharomyces cerevisiae ; Flow cytometry ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract YBR1012 (YBR136w) is an essential gene from Saccharomyces cerevisiae identified during the systematic sequencing of part of the right arm of chromosome II. We previously constructed a conditional allele of YBR1012 based on antisense RNA, by inserting a small fragment of this gene downstream from the inducible UASGAL10-CYC1 promoter. Several other antisense RNA constructions have since been made and their activity tested. The response of the system appears to be very delicate, as the presence or absence of 13 nucleotides of polylinker in the 300 nucleotide antisense transcript can dramatically modify its effectiveness. The most effective antisense RNA construction was used in flow cytometry studies to investigate the role of ybr1012p. The results show that during the antisense RNA block some 80% of the cells are arrested with their DNA unreplicated, suggesting that Ybr1012p is needed for progression through G1 or early S phase.
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    Inactivation of the yeast Sen1 protein affects the localization of nucleolar proteins (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 571-584 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; SEN1 ; tRNA splicing ; RNA interaction ; Nuclear transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mutation in the Saccharomyces cerevisiae SEN1 gene causes accumulation of end-matured, intron-containing pre-tRNAs. Cells containing the thermosensitive sen1-1 mutation exhibit reduced tRNA splicing endonuclease activity. However, Sen1p is not the catalytic subunit of this enzyme. We have used Sen1p-specific antibodies for cell fractionation studies and immunofluorescent microscopy and determined that Sentp is a low abundance protein of about 239 kDa. It localizes to the nucleus with a granular distribution. We verified that a region in SEN1 containing a putative nuclear localization signal sequence (NLS) is necessary for nuclear targeting. Furthermore, we found that inactivation of Sen1p by temperature shift of a strain carrying sen1-1 leads to mislocalization of two nucleolar proteins, Nopt and Ssb1 Possible mechanisms are discussed for several related nuclear functions of Sen1p, including tRNA splicing and the maintenance of a normal crescent-shaped nucleolus.
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    Heat shock protein HSP60 can alleviate the phenotype of mitochondrial RNA-deficient temperature-sensitive mna2 pet mutants (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 56-64 
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    ISSN: 1617-4623
    Keywords: Temperature-sensitive mutants ; Heat shock protein 60 ; Conservative single amino acid substitutions ; Mitochondrial biogenesis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract mna2, which belongs to the class I temperature-sensitive pet mutants that lose mitochondrial (mt)RNA at restrictive temperature, was shown by complementation and sequence determination to correspond to the gene coding for HSP60. Both mna2-1 and mna2-2, the two available alleles of mna2, have conservative single amino acid substitutions in the HSP60 gene. Valine substitutes for an alanine (position 47) in mna2-1, and an isoleucine substitutes for a valine (position 77) in mna2-2. These substitutions result in defects in respiration and in steady-state mtRNA accumulation. Wild-type hsp60 alleviates the mtRNA phenotype completely, while partially relieving the respiratory deficiency.
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    Efficient translation of an SSA1-derived heat-shock mRNA in yeast cells limited for cap-binding protein and eIF-4F (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 619-627 
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    ISSN: 1617-4623
    Keywords: Protein synthesis ; Translation initiation eIF-4E ; cdc33 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Eukaryotic mRNA molecules have a 5′ cap structure that is recognized by the cap-binding component of translation initiation factor eIF-4F during protein synthesis. In the budding yeast Saccharomyces cerevisiae this cap-binding protein is encoded by the CDC33 gene. We report here that decreased global translation initiation in cdc33 mutant cells has virtually no effect on the translation of mRNA from the SSA1 -lacZ chimeric gene, comprised of yeast SSA1 hsp70 gene transcription and translation initiation sequences fused in-frame to the bacterial lacZ gene. When global translation initiation was limited in cdc33 mutant cells, Ssal-LacZ polypeptide synthesis was increased relative to total protein synthesis, and the β-galactosidase activity of the SsaI-LacZ fusion protein was induced to wild-type levels. The normal rate of Ssal-LacZ polypeptide synthesis in mutant cells was maintained by normal levels of SSA1 -lacZ mRNA. Furthermore, in cdc33 mutant cells, the size of polysomes containing SSA1-lacZ mRNA was unaffected, while polysomes containing other specific mRNAs were smaller. Efficient Ssal-LacZ polypeptide synthesis was also seen during eIF-4F limitation produced by disruption of the TIF4631 gene, encoding the large eIF-4F subunit. All of these findings indicate efficient SSA1-lacZ mRNA usage under conditions of globally impaired translation initiation due to eIF-4F limitation.
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    Reactivation of the ATCase domain of the URA2 gene complex: a positive selection method for Ty insertions and chromosomal rearrangements in Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 767-773 
    Details
    ISSN: 1617-4623
    Keywords: Ty retrotransposon ; Duplication ; Saccharomyces cerevisiae ; ATCase ; URA2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic rearrangements such as deletions or duplications of DNA sequences are rarely detected in the yeast Saccharomyces cerevisiae. We have developed a screening system using the URA2 gene coding for the bi-functional CPSase-ATCase (carbamyl phosphate synthetase — aspartate transcarbamylase) to select positively for these kinds of events. Nonsense mutations in the CPSase region cause a complete loss of the ATCase activity because of their strong polar effect. Thirty-seven ATCase+ revertants were isolated from a strain containing three nonsense mutations in the proximal CPSase region. Genetic and structural analysis of the URA2 locus in these strains allowed us to characterize two major classes of revertants. In the first, an entire copy of a Ty transposon was found to be inserted in the CPSase coding domain. This event, which represents a new form of Ty-mediated gene activation was further analysed by mapping the Ty integration site in 26 strains. In a second class of revertants, we observed chromosomal rearrangements and, in particular, duplication of the ATCase region and its integration in a new chromosomal environment in which this sequence becomes active.
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    The C-terminal domain of SIN1 in yeast interacts with a protein that binds the URS1 region of the yeast HO gene (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 774-777 
    Details
    ISSN: 1617-4623
    Keywords: Protein-protein interactions ; Saccharomyces cerevisiae ; SIN1 ; HO
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A protein or protein complex has previously been identified in Saccharomyces cerevisiae which both binds a short DNA sequence in URS1 of HO and interacts with SIN1. SIN1, which has some sequence similarity to mammalian HMG1, is an abundant chromatin protein in yeast and is thought to participate in the transcriptional repression of a specific family of genes. SIN1 binds DNA weakly, though it has no DNA binding specificity. Here we address the nature of the interaction between SIN1 and the specific DNA binding protein(s) to HO DNA. We show that the isolated C-terminal region of SIN1 can interact in vitro with the DNA binding protein, causing a supershift in a gel mobility shift assay. Interestingly, inclusion of the region in SIN1 which contains two acidic sequences, precludes the binding of recombinant protein to the DNA/protein complex.
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    The glucose repression and RAS-cAMP signal transduction pathways of Saccharomyces cerevisiae each affect RNA processing and the synthesis of a reporter protein (1995)
    Springer
    Molecular genetics and genomics 247 (1995), S. 48-54 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; RNA processing ; Signal transduction ; Glucose repression ; RNA1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previously we reported that mutations in the Saccharomyces cerevisiae REG1 gene encoding a negative regulator of glucose-repressible genes, suppress the RNA processing defects and temperature-sensitive growth of rna1-1 and prp cells. This result and the fact that growth on non-glucose carbon sources also suppresses rna1-1 led us to propose that RNA processing and export of RNA from the nucleus are responsive to carbon source regulation. To understand how carbon source affects these processes, we used p70, an antigen regulated by REGI and by glucose availability, as a reporter. We found that the response of p70 to glucose availability is mediated by both the SNFI-SSN6-dependent glucose repression and the RAS-cAMP pathways. These results led us to test whether the RAS-cAMP pathway interacts with RNA1. We found that suppression of rnal-1 appears to be mediated, at least in part, by the RAS-cAMP pathway.
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    URL: http://dx.doi.org/10.1007/BF00425820
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    The role ofSaccharomyces cerevisiae Cdc40p in DNA replication and mitotic spindle formation and/or maintenance (1995)
    Springer
    Molecular genetics and genomics 247 (1995), S. 123-136 
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    ISSN: 1617-4623
    Keywords: CDC40 ; DNA replication ; Mitotic spindle assembly ; cyclins ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Successful progression through the cell cycle requires the coupling of mitotic spindle formation to DNA replication. In this report we present evidence suggesting that, inSaccharomyces cerevisiae, theCDC40 gene product is required to regulate both DNA replication and mitotic spindle formation. The deduced amino acid sequence ofCDC40 (455 amino acids) contains four copies of a β-transducin-like repeat. Cdc40p is essential only at elevated temperatures, as a complete deletion or a truncated protein (deletion of the C-terminal 217 amino acids in thecdc40-1 allele) results in normal vegetative growth at 23°C, and cell cycle arrest at 36°C. In the mitotic cell cycle Cdc40p is apparently required for at least two steps: (1) for entry into S phase (neither DNA synthesis, nor mitotic spindle formation occurs at 36°C and (2) for completion of S-phase (cdc40::LEU2 cells cannot complete the cell cycle when returned to the permissive temperature in the presence of hydroxyurea). The role of Cdc40p as a regulatory protein linking DNA synthesis, spindle assembly/maintenance, and maturation promoting factor (MPF) activity is discussed.
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    URL: http://dx.doi.org/10.1007/BF00705642
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    Isolation and characterization of temperature-sensitiveplc1 mutants of the yeastSaccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 247 (1995), S. 148-156 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Phosphoinositide-specific phospholipase C ; Temperature-sensitive mutant ; PLC1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract ThePLC1 gene of the yeastSaccharomyces cerevisiae has been discovered to encode a homolog of mammalian phosphoinositide-specific phospholipase C (PLC). Five temperature-sensitiveplc1 mutants were isolated by in vitro mutagenesis with subsequent plasmid shuffling. All of the amino acid substitutions that caused a temperature-sensitive growth phenotype were located in the X or the Y region, both of which are conserved among PLC isoenzymes. The PLC activity of all products of mutantplc1 genes was dramatically lower than that of the wild-type product, indicating that PLC activity itself is important for cell growth. At the restrictive temperature,plc1 mutant cells ceased growth at random times during the cell cycle, a result that suggests thatPLC1 is required at several or all stages of the cell cycle.
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    URL: http://dx.doi.org/10.1007/BF00705644
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    Constitutive activation of the Saccharomyces cerevislae mating response pathway by a MAP kinase kinase from Candida albicans (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 609-621 
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    ISSN: 1617-4623
    Keywords: MAP kinase kinase ; Ste7 ; Saccharomyces cerevisiae ; Candida albicans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The HST7 gene of Candida albicans encodes a protein with structural similarity to MAP kinase kinases. Expression of this gene in Saccharomyces cerevisiae complements disruption of the Ste7 MAP kinase kinase required for both mating in haploid cells and pseudohyphal growth in diploids. However, Hst7 expression does not complement loss of either the Pbs2 (Hog4) MAP kinase kinase required for response to high osmolarity, or loss of the Mkk1 and Mkk2 MAP kinase kinases required for proper cell wall biosynthesis. Intriguingly, HST7 acts as a hyperactive allele of STE7; expression of Hst7 activates the mating pathway even in the absence of upstream signaling components including the Ste7 regulator Ste11, elevates the basal level of the pheromone-inducible FUS1 gene, and amplifies the pseudohyphal growth response in diploid cells. Thus Hst7 appears to be at least partially independent of upstream activators or regulators, but selective in its activity on downstream target MAP kinases. Creation of Hst7/Ste7 hybrid proteins revealed that the C-terminal two-thirds of Hst7, which contains the protein kinase domain, is sufficient to confer this partial independence of upstream activators.
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    URL: http://dx.doi.org/10.1007/BF00418030
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    Alternative photophosphorylation, inorganic pyrophosphate synthase and inorganic pyrophosphate (1995)
    Springer
    Photosynthesis research 46 (1995), S. 87-91 
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    ISSN: 1573-5079
    Keywords: bioenergetics ; photosynthesis ; chromatophores ; energy coupling ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This minireview in memory of Daniel I. Arnon, pioneer in photosynthesis research, concerns properties of the first and still only known alternative photophosphorylation system, with respect to the primary phosphorylated end product formed. The alternative to adenosine triphosphate (ATP), inorganic pyrophosphate (PPi), was produced in light, in chromatophores from the photosynthetic bacterium Rhodospirillum rubrum, when no adenosine diphosphate (ADP) had been added to the reaction mixture (Baltscheffsky H et al. (1966) Science 153: 1120–1122). This production of PPi and its capability to drive energy requiring reactions depend on the activity of a membrane bound inorganic pyrophosphatase (PPase) (Baltscheffsky M et al. (1966) Brookhaven Symposia in Biology, No. 19, pp 246–253); (Baltscheffsky M (1967) Nature 216: 241–243), which pumps protons (Moyle J et al. (1972) FEBS Lett 23: 233–236). Both enzyme and substrate in the PPase (PPi synthase) are much less complex than in the case of the corresponding adenosine triphosphatase (ATPase, ATP synthase). Whereas an artificially induced proton gradient alone can drive the synthesis of PPi, both a proton gradient and a membrane potential are required for obtaining ATP. The photobacterial, integrally membrane bound PPi synthase shows immunological cross reaction with membrane bound PPases from plant vacuoles (Nore BF et al. (1991) Biochem Biophys Res Commun 181: 962–967). With antibodies against the purified PPi synthase clones of its gene have been obtained and are currently being sequenced. Further structural information about the PPi synthase may serve to elucidate also fundamental mechanisms of electron transport coupled phosphorylation. The existence of the PPi synthase is in line with the assumption that PPi may have preceded ATP as energy carrier between energy yielding and energy requiring reactions.
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    URL: http://dx.doi.org/10.1007/BF00020419
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    Variation and phylogeny of the triploid cultivated potatoSolanum chaucha Juz. et Buk. based on RAPD and isozyme markers (1995)
    Springer
    Genetic resources and crop evolution 42 (1995), S. 373-386 
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    ISSN: 1573-5109
    Keywords: evolution ; genetic distance ; isozymes ; RAM ; Solanum chaucha ; varietal classification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Ninety four accessions of the cultivated triploid potatoS. chaucha were analyzed and classified in genotypic groups using 9 isozyme loci and RAPD markers disclosed by 20 arbitrary 10-mer primers. Eight isozyme loci out of nine were polymorphic. A total of 22 allozymes were analyzed but none of them were specific for any genotypic group. About half (52%) of the 102 RAPD markers scored, were polymorphic, all of them showing polymorphism among groups and rarely within groups. Eighteen RAPD markers were specific for certain genotypes. The isozyme markers showed a certain amount of intra group variation which made classification less reliable than with RAPD markers. A total of 10 triploid genetic groups were discriminated using both techniques together. A single primer was found to be sufficient to distinguish all 10 groups. All varieties of a single group are considered to have been derived from the same cross and then clonally propagated, even though there is a high amount of morphological variation within a single genotypic group due probably to somatic mutations. RAPD markers have been shown to be more reliable in the classification of triploid potato varieties than other genetic markers like isozymes, proteins and morphological traits.
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    URL: http://dx.doi.org/10.1007/BF02432142
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    Sequence evolution of theGpdh gene in theDrosophila virilis species group (1995)
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    Genetica 96 (1995), S. 293-302 
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    ISSN: 1573-6857
    Keywords: D. virilis phylad ; evolution ; Gpdh gene ; nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleotide sequence of theGpdh gene from six taxa,D. virilis, D. lummei, D. novamexicana, D. a. americana, D. a. texana andD. ezoana, belonging to thevirilis species group was determined to examine details of evolutionary change in the structure of theGpdh gene. TheGpdh gene is comprised of one 5′ non-translated region, eight exons, seven introns and three 3′ non-translated regions. Exon/intron organization was identical in all the species examined, but different from that of mammals. Interspecific nucleotide divergence in the entireGpdh gene followed the common pattern: it was low in the exon, high in the intron and intermediate in the non-translated regions. The degree of nucleotide divergence differed within these regions, suggesting that selection exerts constraints differentially on nucleotide change of theGpdh gene. A phylogenetic tree of thevirilis phylad constructed from nucleotide variation of total sequence was consistent with those obtained from other data.
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    Dormancy in the Copepoda — an overview (1995)
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    Hydrobiologia 306 (1995), S. 199-211 
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    ISSN: 1573-5117
    Keywords: Diapause ; ecology ; evolution ; life history ; marine ; freshwater ; Copepoda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dormancy affects copepods in their anatomy, physiology, genetics, population biology, community ecology, evolution and local and geographic distribution. It is known from freeliving representatives of three copepod taxa, namely the Harpacticoida, Cyclopoida and Calanoida. Species showing dormancy occur in various realms and habitats, both freshwater and marine, being benthic, planktic or ice-dwelling. Depending on the taxon, dormancy occurs at various times of the year, prevailing in higher and temperate latitudes. Copepod dormancy is expressed in various ontogenetic stages, such as resting eggs, arrested larval development, juvenile and adult encystment, or arrested development of nonencysted copepodids or adults. Ecologically, dormancy is an energy saving trait, allowing the individual to bridge periods of environmental harshness. Adverse environmental conditions could be abiotic (e.g. desiccation, temperature, oxygen availability) or biotic in nature (e.g. food availability, predation). Diapause s. str. is initiated, maintained and terminated by triggering factors (e.g. photoperiod, temperature, chemical cues, population density/physiological factors). The dormant state and emergence patterns directly affect reproduction, population dynamics, community composition, coexistence and distribution of copepods, as well as the phenology of their predators and living food items. Populations having dormancy, in most cases belong to and affect communities of two realms: the water column and the bottom. Dormant stages may provide means for dispersal as well as for staying in special localities. The variability of dormancy permits flexible and complex life histories. Dormancy is subjected to and on the other hand affects copepod evolution.
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    Continuous production of non-alcohol beer by immobilized yeast at low temperature (1995)
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    Journal of industrial microbiology and biotechnology 14 (1995), S. 495-501 
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    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Non-alcohol beer ; Wort ; Immobilization ; DEAE-cellulose carrier ; Low temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A system for production of non-alcohol beer is described. A limited fermentation is carried out with immobilized cells ofSaccharomyces cerevisiae in a packed bed reactor. In the reactor, combined stress factors such as low temperature (2–4°C) and anaerobic conditions limit cell metabolism. Of the available sugars only a small amount of glucose is metabolized, resulting in low concentrations of ethanol (〈0.08%). The absence of oxygen affects the redox balance of the yeast cell, and thus stimulates formation of esters and higher alcohols. Products are formed by reduction of wort aldehydes, as well as reduction of intracellular metabolites. Despite the stress conditions, biomass increases during prolonged production periods. In batch experiments,S. cerevisiae strain W34 grows at low temperatures and a mininum growth temperature of −2 °C was found, indicating that a further reduction of temperature during production will not inhibit growth. The characteristics of the system allow its use in very different applications. Potential applications of the immobilized system are discussed.
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    Adaptive on-line simulation of bioreactors: Fermentation monitoring and modeling system (1995)
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    Journal of industrial microbiology and biotechnology 14 (1995), S. 403-411 
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    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Process control ; State estimation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary In order to study and control fermentation processes, indirect on-line measurements and mathematical models can be used. Here an on-line model for fermentation processes is presented. The model is based on atom and partial mass balances as well as on stability equations for the protolytes. The model is given an adaptive form by including transport equations for mass transfer and expressions for the fermentation kinetics. The state of the process can be estimated on-line using the balance component of the model completed with measurement equations for the input and the output flows of the process. Adaptivity is realized by means of on-line estimation of the parameters in the transport and kinetic expressions using recursive regression analysis. On-line estimation of the kinetic and mass transfer parameters makes model-based predictions possible and enables intelligent process control while facilitating testing of the validity of the measurement variables. A practical MS-Windows 3.1 model implementation called FMMS—Fermentation Monitoring and Modeling System is shown. The system makes it easy to configure the operating conditions for a run. It uses Windows dialogs for all set-ups, model configuration parameters, elemental compositions, on-line measurement devices and signal conditioning. Advanced on-line data analysis makes it possible to plot variables against each other for easy comparison. FMMS keeps track of over 100 variables per run. These variables are either measured or estimated by the model. Assay results can also be entered and plotted during fermentation. Thus the model can be verified almost instantly. Historical fermentation runs can be re-analyzed in simulation mode. This makes it possible to examine different signal conditining filters as well as the sensitivity of the model. Combined, the data analysis and the simulation mode make it easy to test and develop model theories and new ideas.
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    Chemical and cytological changes during the autolysis of yeasts (1995)
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    Journal of industrial microbiology and biotechnology 14 (1995), S. 440-450 
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    ISSN: 1476-5535
    Keywords: Yeasts ; Autolysis ; Saccharomyces cerevisiae ; Kloeckera apiculata ; Candida stellata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Cell suspensions ofSacharomyces cerevisiae, Kloeckera apiculata andCandida stellata were autolyzed in phosphate buffer, pH 4.5, for up to 10 days. Cell dry weights decreased by 25–35% after 10 days. Based on initial cell dry weight, the soluble autolysate consisted of: carbohydrate (principally polysaccharide) 3–7%; organic acids 3–6%; protein 12–13%; free amino acids 8–12%; nucleic acid products 3–5%; and lipids 1–12%. The main organic acids in autolysates were propionic, succinic and acetic and the main amino acids were phenylalanine, glutamic acid, leucine, alanine and arginine. Approximately 85–90% of cellular RNA and 25–40% of cellular DNA were degraded during autolysis. Both neutral lipid and phospholipid components were degraded, with neutral lipids but not phospholipids being found in autolysates. Scanning and transmission electron micrographs showed retention of cell wall structure and shape during autolysis, but there was extensive intracellular disorganization withinS. cerevisiae andC. stellata. There were differences in the autolytic behavior ofK. apiculata compared withS. cerevisiae andC. stellata.
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    Molecular cloning, expression and evaluation of phosphohydrolases for phytate-degrading activity (1995)
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    Journal of industrial microbiology and biotechnology 14 (1995), S. 396-402 
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    ISSN: 1476-5535
    Keywords: Acid phosphatase ; Phytase ; Aspergillus ; Saccharomyces cerevisiae ; Phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Four acid phosphatase (phosphomonoesterase E.C.3.1.3.2) genes, werecloned by polymerase chain reaction (PCR). These were pho3, pho5 and pho11 fromSaccharomyces cerevisiae and the gene for a phosphate-respressible acid phosphatase fromAspergillus niger. The individual genes were subcloned into anA. oryzae expression vector downstream from a starch-inducible α-amylase promoter and the resulting expression constructs were transformed into a mutant strain ofA. oryzae, AO7. Southern hybridization analysis confirmed that the acid phosphatase genes had been integrated into the host genome with estimates of integrated copy numbers ranging from 2 to 20 for individual transformants. Northern hybridization analysis of total RNA from individual transformants revealed the presence of a single transcript of the expected size of 1.8 kb. Production of recombinant protein was induced by the addition of 30 g L−1 of soluble starch in the fermentationmedia. Active acid phosphatases, not present in control cultures, were detected in the supernatant fractions of transformant cultures by acid phosphatase activity staining of non-denaturing polyacrylamide gels. The ability of the recombinant acid phosphatases to hydrolyze phytate was assessed by referenced phytase (myoinositol hexakisphosphate phosphohydrolase E.C. 3.1.3.8) activity assay procedures. A two- to six-fold increase in phytase activity was measured in transformants compared to control, untransformedA. oryzae. Sufficient quantities ofA. niger and pho5 recombinant acid phosphatases were generated from large-scale fermentations to assess the efficacy of these enzymes as phytate-degrading enzymes when included in poultry diets. Data indicated an increase in available phosphorus of 1 g kg−1 obtained with yeast acid phosphatase andA. niger acid phosphatase representing 40% utilization of unavailable dietary P compared to 48% utilization for commercial phytase.
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    Industrial yeast strain improvement: construction of a highly flocculent yeast with a killer character by protoplast fusion (1995)
    Springer
    Journal of industrial microbiology and biotechnology 15 (1995), S. 94-102 
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    ISSN: 1476-5535
    Keywords: protoplast fusion ; killer character ; flocculence ; Saccharomyces cerevisiae ; industrial yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Conditions were optimized for rapid release and improved regeneration of protoplasts ofSaccharomyces cerevisiae NCIM 3458. Rapid protoplast release was also obtained with representatives of several other yeast genera under the modified conditions of treatment. The application of the procedure in construction of a highly flocculentSaccharomyces cerevisiae with a killer character is described. Fusion was effected between UV-killed protoplasts ofS. cerevisiae NCIM 3578 with a killer character and live protoplasts of the highly flocculentS. cerevisiae NCIM 3528 in the presence of polyethylene glycol (PEG) 6000. Fusants were selected using benomyl resistance as marker, the killer toxin producer rather than the highly flocculent yeast being resistant to the fungicide at a concentration of 100 μg ml−1. Fusants were also characterized by their DNA contents, capacity for ethanolic fermentation of molasses sugar and levels of invertase, alcohol dehydrogenase and pyruvate decarboxylase activities.
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    Flocculation of industrial and laboratory strains ofSaccharomyces cerevisiae (1995)
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    Journal of industrial microbiology and biotechnology 14 (1995), S. 461-466 
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    ISSN: 1476-5535
    Keywords: Flocculation ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A comparative study has been made of different laboratory and industrial wild-type strains ofSaccharomyces cerevisiae in relation to their flocculation behavior. All strains were inhibited by mannose and only one by maltose. In regard to the stability of these characters in the presence of proteases and high salt concentrations, a relevant degree of variation was found among the strains. This was to such an extent that it did not allow their inclusion in the Flol or NewFlo phenotypes. Genetic characterization of one wild-type strain revealed that the flocculation-governing gene was allelic toFLO1 found in genetic strains.
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    Evaluation of image analysis and laser granulometry for microbial cell sizing (1995)
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    Antonie van Leeuwenhoek 67 (1995), S. 139-149 
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    ISSN: 1572-9699
    Keywords: image analysis ; laser granulometry ; cell size ; cell morphology ; Zymomonas mobilis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A direct cell size measurement technique and an image analysis based sizing method were developed. The former consisted of a manual size measurement of the two-dimensional cell images on a video screen, with automatic data recording. This method was chosen as the reference. The latter, a semiautomatic method took advantage of a commercial computer program designed for image processing and particle morphology analysis. It gave average and median size values which were compatible with the manual method. However, the performance of these time consuming methods is limited. Hence, the laser granulometry technique, intrinsically far more powerful while capable of analysing millions of sample objects in a short time delay, was applied. The comparison revealed that this method gives too low size values, particularly in disagreement with the known dimensions of the bacterial (Zymomonas mobilis) cells. A size correction method was developed to realign the granulometry results ofZ. mobilis cell samples with those of the direct manual measurement method.
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    Effects of growth conditions on mitochondrial morphology inSaccharomyces cerevisiae (1995)
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    Antonie van Leeuwenhoek 67 (1995), S. 243-253 
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    ISSN: 1572-9699
    Keywords: mitochondria ; morphology ; Saccharomyces cerevisiae ; vital staining ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effects of growth conditions on mitochondrial morphology were studied in livingSaccharomyces cerevisiae cells by vital staining with the fluorescent dye dimethyl-aminostyryl-methylpyridinium iodine (DASPMI), fluorescence microscopy, and confocal-scanning laser microscopy. Cells from respiratory, ethanol-grown batch cultures contained a large number of small mitochondria. Conversely, cells from glucose-grown batch cultures, in which metabolism was respiro-fermentative, contained small numbers of large, branched mitochondria. These changes did not significantly affect the fraction of the cellular volume occupied by the mitochondria. Similar differences in mitochondrial morphology were observed in glucose-limited chemostat cultures. In aerobic chemostat cultures, glucose metabolism was strictly respiratory and cells contained a large number of small mitochondria. Anaerobic, fermentative chemostat cultivation resulted in the large, branched mitochondrial structures also seen in glucose-grown batch cultures. Upon aeration of a previously anaerobic chemostat culture, the maximum respiratory capacity increased from 10 to 70 µmole.min−1.g weight−1 within 10 h. This transition resulted in drastic changes of mitochondrial number, morphology and, consequently, mitochondrial surface area. These changes continued for several hours after the respiratory capacity had reached its maximum. Cyanide-insensitive oxygen consumption contributed ca. 50% of the total respiratory capacity in anaerobic cultures, but was virtually absent in aerobic cultures. The response of aerobic cultures to oxygen deprivation was qualitatively the reverse of the response of anaerobic cultures to aeration. The results indicate that mitochondrial morphology inS. cerevisiae is closely linked to the metabolic activity of this yeast: conditions that result in repression of respiratory enzymes generally lead to the mitochondrial morphology observed in anaerobically grown, fermenting cells.
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    Molecular evolution in bacteria (1995)
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    Antonie van Leeuwenhoek 67 (1995), S. 315-324 
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    ISSN: 1572-9699
    Keywords: bacteria ; ecology ; evolution ; metabolism ; microbiology ; molecular biology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Recent advances in microbiology and molecular biology have a unifying influence on our understanding of genetic diversity/similarity and evolutionary relationships in microorganisms. This article attempts to unify information from diverse areas such as microbiology, molecular biology, microbial physiology, clay crystal genes, metals-microbe-clay interactions and bacterial DNA restriction-modification systems (R-M) as they may apply to molecular evolution of bacteria. The possibility is discussed that the first informational molecules may have been catalytic RNA (micro-assembler) not DNA (now the master copy) and these first micro-assemblers may have been precursors of ribosomes.
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    Phylogenesis of fission yeasts. Contradictions surrounding the origin of a century old genus (1995)
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    Antonie van Leeuwenhoek 68 (1995), S. 119-149 
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    ISSN: 1572-9699
    Keywords: Schizosaccharomyces ; sequence comparison ; evolution ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phylogenesis of fungi is controversial due to their simple morphology and poor fossilization. Traditional classification supported by morphological studies and physiological traits placed the fission yeasts in one group with ascomycetous yeasts. The rRNA sequence comparisions, however, revealed an enormous evolutionary gap betweenSaccharomyces andSchizosaccharomyces. As shown in this review, the protein sequences also show a large gap which is almost as large as that separatingSchizosaccharomyces from higher animals. Since the two yeasts share features (both cytological and molecular) in common which are also characteristic of ascomycetous fungi, their separation must have taken place later than the sequence differences may suggest. Possible reasons for the paradox are discussed. The sequence data also suggest a slower evolutionary rate in theSchizosaccharomyces lineage than in theSaccharomyces branch. In the fission yeast lineage two ramifications can be supposed. FirstS. japonicus (Hasegawaea japonica) branched off, thenS. octosporus (Octosporomyces octosporus) separated fromS. pombe.
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    High molecular weight precursors of glucans inSaccharomyces cerevisiae (1995)
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    Antonie van Leeuwenhoek 68 (1995), S. 231-235 
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    ISSN: 1572-9699
    Keywords: beta glucan ; glucan synthetase ; protein acceptor ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nascent β-1,3 glucan synthesized by mixed membrane fractions fromSaccharomyces cerevisiae was solubilized by extraction with hot SDS or urea. Nature of the material was analyzed by electrophoresis and gel filtration. As determined by gel filtration, Mr of synthesized glucans exceeded 1,500 kDa, but was below 20,000 kDa. This nascent material served as an acceptor for further glucose transfer reactions, giving rise to glucan molecules over 20,000 kDa. It is suggested that the high Mr precursor components represent protein-bound glucan molecules in transit to the cell surface.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00871820
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  • 95
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    Qualitative mathematical discussion of different evolutionary states in water transport systems of plants (1995)
    Springer
    Journal of biological physics 21 (1995), S. 241-264 
    Details
    ISSN: 1573-0689
    Keywords: Synergetics ; dynamical systems ; water transport ; evolution ; stele
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract We shall present several qualitative mathematical models to describe the early evolution of water transport systems in plants. To perform this in a systematic way we apply methods which have been developed in phenomenological synergetics. These methods rest on the fact that it becomes possible to describe the macroscopic behavior of a complex system by a set of control and order parameters when they are suitably identified. Our presentation is addressed to community with interdisciplinary interests.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00700627
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  • 96
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    Variation and phylogeny of the ribosomal DNA unit types and 5 S DNA inPetunia Jussieu (1995)
    Springer
    Genetic resources and crop evolution 42 (1995), S. 311-325 
    Details
    ISSN: 1573-5109
    Keywords: evolution ; 5 S DNA ; Petunia ; ribosomal DNA RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Seven wildPetunia species including 2n = 18 species (P. parviflora Jussieu,P. linearis Hook.) and those with 2n = 14 (P. parodii Steene,P. axillaris Lam.,P. integrifolia Hook.,P. inflata R.E. Fries,P. violacea Lindl.) and tenPetunia hybrida horticultural lines were compared for polymorphisms in rDNA genes using the four restriction enzymesEcoRI,BamHI,HindIII andXhoI. All the unit types found in the lines pre-existed in the wild forms. There are two different sizes of either 11.45 or 11.6 kb./The 2n = 18 species are closely related to the 2n = 14 species, thus making thePetunia genus homogeneous. Moreover, it is likely thatP. hybrida lines originated in several kinds of crosses between these species. We constructed a dendrogram for all the 15 rDNA unit types found. Two main branches of the tree result from the presence or the absence ofHindIII sites. The main branch is divided according to variability at theEcoRI andBamHI sites. Taking into account the existence of several loci which carry one unit type only, we consider whether or not exchanges might occur between loci. Lines carrying two unit types and lines carrying three unit types support such a hypothesis.XhoI andBamHI fragments enable us to distinguish two types of 5S DNA corresponding to 2n = 18 and 2n = 14 species, respectively.P. hybrida lines and each 2n = 14 wild species carry one of the types only, that corresponds to one 5S DNA locus. The most parsimonious phylogenetic trees whatever the species chosen as the outgroup, do not fit with our knowledge ofPetunia and with taxonomy. This is likely because only few loci formed the basis of these phylogenetic constructions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02432135
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  • 97
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    Discovery of 2n gametes in tetraploid oat Avena vaviloviana (1995)
    Springer
    Euphytica 81 (1995), S. 1-6 
    Details
    ISSN: 1573-5060
    Keywords: Avena ; 2n gametes ; binucleate cell ; evolution ; sexual polyploidization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sexual polyploidization via the action of 2n gametes (gametes with the sporophytic chromosome number) has been identified as the most important evolutionary mode of polyploidization among plant genera. This study was conducted to determine whether 2n gametes are present in the tetraploid level of the genus Avena (2n=4×=28) Twenty tetraploid Avena lines, representing four species and one interspecific hybrid, were screened for pollen grain size in order to differentiate between n and 2n pollen. Avena vaviloviana (Malz.) Mordv. line PI 412767 was observed to contain large pollen grains at a 1.0% frequency. Cytogenetic analyses of pollen mother cells of PI 412767 revealed cells with double the normal chromosome number (i.e., 56 chromosomes at metaphase I and anaphase I). The mode of chromosome doubling was found to be failure of cell wall formation during the last mitotic division that preceded meiosis. The resulting binucleate cells underwent normal meiotic divisions and formed pollen grains with 28 chromosomes. Based on the formation and function of 2n gametes, three models involving diploid and tetraploid oat lines are proposed to describe possible evolutionary pathways for hexaploid oats. If stable synthetic hexaploid oat lines could be developed by utilizing 2n gametes from diploid and tetraploid oat species through bilateral sexual polyploidization, the resulting hexaploids could be used in breeding programs for transferring genes from diploids and tetraploids to cultivated hexaploids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00022452
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  • 98
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    Evolutionary aspects of gonadotropin-releasing hormone and its receptor (1995)
    Springer
    Cellular and molecular neurobiology 15 (1995), S. 5-23 
    Details
    ISSN: 1573-6830
    Keywords: evolution ; reproduction ; gonadotropin-releasing hormone (GnRH) structure ; GnRH function ; GnRH receptor structure ; GnRH receptor function
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Gonadotropin-releasing hormone (GnRH) was originally isolated as a hypothalamic peptide hormone that regulates the reproductive system by stimulating the release of gonadotropins from the anterior pituitary. However, during evolution the peptide was subject to gene duplication and structural changes, and multiple molecular forms have evolved. 2. Eight variants of GnRH are known, and at least two different forms are expressed in species from all vertebrate classes: chicken GnRH II and a second, unique, GnRH isoform. 3. The peptide has been recruited during evolution for diverse regulatory functions: as a neurotransmitter in the central and sympathetic nervous systems, as a paracrine regulator in the gonads and placenta, and as an autocrine regulator in tumor cells. 4. Evidence suggests that in most species the early-evolved and highly conserved chicken GnRH II has a neurotransmitter function, while the second form, which varies across classes, has a physiologic role in regulating gonadotropin release. 5. We review here evolutionary aspects of the family of GnRH peptides and their receptors.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02069556
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  • 99
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    Changes in the intracellular concentrations of the adenosine phosphates and nicotinamide adenine dinucleotides ofSaccharomyces cerevisiae during batch fermentation (1995)
    Springer
    World journal of microbiology and biotechnology 11 (1995), S. 196-201 
    Details
    ISSN: 1573-0972
    Keywords: Adenosine phosphates ; fermentation ; flor-veil-forming yeast ; nicotinamide adenine dinucleotides ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Significant changes in the intracellular concentrations of adenosine phosphates and nicotinamide adenine dinucleotides were observed during fermentation of grape must by three different strains ofSaccharomyces cerevisiae: S. cerevisiae var.cerevisiae, a typical fermentative yeast strain and two flor-veil-forming strains,S. cerevisiae var.bayanus andS. cerevisiae var.capensis. The intracellular concentration of ATP was always higher inS. cerevisiae var.cerevisiae than in the flor-veil-forming strains. NAD+ and NADP+ concentrations decreased at faster rates in the flor-veil-forming yeasts than in the other yeast but NADH concentration was the same in all yeasts for the first 10 days of fermentation. NADPH concentration was always lower inS. cerevisiae var.cerevisiae than in the other yeasts and this yeast also showed higher rates of growth and fermentation during the early stages of the fermentation and the presence of non-viable cells at the end of fermentation. In contrast, the flor-veil-forming strains maintained growth and fermentation capabilities for a relatively long time and viable cells were present throughout the entire fermentation process (31 days).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00704648
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  • 100
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    A gene-culture model of human handedness (1995)
    Springer
    Behavior genetics 25 (1995), S. 433-445 
    Details
    ISSN: 1573-3297
    Keywords: Handedness ; asymmetry ; genetic ; cultural transmission ; mathematical model ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract A model of handedness incorporating both genetic and cultural processes is proposed, based on an evolutionary analysis, and maximum-likelihood estimates of its parameters are generated. This model has the characteristics that (i) no genetic variation underlies variation in handedness, and (ii) variation in handedness among humans is the results of a combination of cultural and developmental factors, but (iii) a genetic influence remains since handedness is a facultative trait. The model fits the data from 17 studies of handedness in families and 14 studies of handedness in monozygotic and dizygotic twins. This model has the additional advantages that it can explain why monozygotic and dizygotic twins and siblings have similar concordance rates, and no hypothetical selection regimes are required to explain the persistence of left handedness.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02253372
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