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  • Yeast  (34)
  • Springer  (34)
  • American Chemical Society
  • American Institute of Physics (AIP)
  • 1985-1989  (34)
  • 1935-1939
  • 1988  (34)
Collection
Keywords
Publisher
  • Springer  (34)
  • American Chemical Society
  • American Institute of Physics (AIP)
  • Wiley-Blackwell  (1)
Years
  • 1985-1989  (34)
  • 1935-1939
Year
  • 1
    Electronic Resource
    Electronic Resource
    Thermotolerant single cell protein production byKluyveromyces marxianus var.marxianus (1988)
    Springer
    Journal of industrial microbiology and biotechnology 3 (1988), S. 9-14 
    Details
    ISSN: 1476-5535
    Keywords: Single cell protein ; Sucrose ; Yeast ; Thermotolerance ; Fermentation ; Kluyveromyces marxianus var.marxianus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Amino acid analyses were undertaken on single cell protein (SCP) produced by thermotolerant strains ofKluyveromyces marxianus var.marxianus grown on sugar cane molasses at 40°C. The maximum conversion of available sugars to biomass at 45°C was only 10.8% (g dry wt.·g−1 total sugars). The amino acid composition of the SCP did not differ markedly from that reported for other yeast species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569436
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  • 2
    Electronic Resource
    Electronic Resource
    Immunological homologies between yeast ribosomal protein L2 and rat liver ribosomal proteins L4 and L24 (1988)
    Springer
    Current genetics 13 (1988), S. 235-239 
    Details
    ISSN: 1432-0983
    Keywords: Ribosomal protein ; Immunological homology ; Yeast ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Polyclonal antibodies raised against ribosomal protein (r-protein) L2 of Schizosaccharomyces pombe were used to check for cross-reaktions with total r-proteins of rat liver. Using this procedure, the rat liver r-proteins, L4 and L24, were identified as being immunologically related to yeast L2. In addtional, homologies between rat liver L4 and L24 were detected. The possible implications for the regulation of r-protein synthesis are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00387769
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  • 3
    Electronic Resource
    Electronic Resource
    Plasmid associations with residual nuclear structures in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 291-297 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Nuclear matrix ; Plasmid stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Acentric yeast plasmids are mitotically unstable, apparently because they cannot freely diffuse after replicating and therefore are not included in the daughter nucleus. This behavior could result if plasmids remain attached to structural elements of the nucleus after replicating. Since DNA replication is believed to take place on the nuclear matrix, we tested whether there was a correlation between the mitotic stability of a given plasmid and the extent to which it was found associated with residual nuclear structures. Residual nuclei were prepared from yeast nuclei by extraction with either high salt, 2 M NaCl, or low salt, 10 mM lithium diiodosalicylate (LIS). Hybridization analysis was used to estimate the fraction of plasmid molecules remaining after nuclei were extracted. We examined the extent of matrix association of three ARSI plasmids, Trpl-RI circle (1.45 kb), YRp7 (5.7 kb) and pXBAT (45.1 kb) with mitotic loss rates ranging from 3–25%. In addition we examined the matrix binding of the endogenous 2 μm plasmid and the 2 μm-derived YEp 13 which is relatively stable in the presence of 2 μm and less stable in cir° strains. Among the ARS1 plasmids we observed a negative correlation between stability and matrix association, consistent with models in which binding to the nuclear matrix prevents passive segregation of ARS1 plasmid molecules. No such correlation was observed among the 2 μn plasmids. Among all plasmids examined there is a positive correlation between size and matrix association.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00424422
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  • 4
    Electronic Resource
    Electronic Resource
    Cloning of the orotidine 5′-phosphate decarboxylase (ODC) gene of Schwanniomyces occidentalis by complementation of the ura3 mutation in S. cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 29-35 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Cloning ; ODC ; Complementation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A DNA fragment containing the gene encoding orotidine 5′-phosphate decarboxylase (ODC) from the yeast, Schwanniomyces occidentalis (formerly castellii) has been isolated from a genomic library constructed in the S. cerevisiae expression vector, pYcDE8. A recombinant plasmid, p2-lA, containing a 2.47 kb insert was shown to complement the ura3-52 mutation of several strains of S. cerevisiae. This DNA insert was shown to be from Schwanniomyces occidentalis by Southern hybridization analysis. A restriction enzyme cleavage map of the insert has been derived and the ODC gene localized to a 1.1 kb region by deletion analysis. In addition, we have demonstrated that expression of ODC is not dependent on the ADHI promoter carried on pYcDE8. This is the first report of the cloning of a gene from a member of the genus Schwanniomyces.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365753
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  • 5
    Electronic Resource
    Electronic Resource
    The glucose- and ethanol-dependent regulation of PDC1 from Saccharomyces cerevisiae are controlled by two distinct promoter regions (1988)
    Springer
    Current genetics 14 (1988), S. 337-344 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Gene regulation ; Saccharomyces cerevisiae ; PDCI promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 870 by promoter fragment of the PDC1 gene that includes the carbon source dependent regulatory regions was investigated using 5′ and 3′ promoter deletions. The results indicate that glucose and ethanol regulation of PDC1 transcription are independently controlled by distinct cis-acting regions. The consensus sequence AAATCGATA may play a role in this regulation, while the sequence (ATCA)AACCT may be important in transcription initiation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419991
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  • 6
    Electronic Resource
    Electronic Resource
    Multiple elements regulate expression of the cell cycle-regulated thymidylate synthase gene of Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 363-373 
    Details
    ISSN: 1432-0983
    Keywords: Gene regulation ; Cell cycle ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Expression of the thymidylate synthase gene (TMPI) of Saccharomyces cerevisiae increases during the late G1 phase of the cell cycle. Using a series of gene fusions, which have placed the Escherichia coli lacZ gene under transcriptional and translational control of different portions of the TMPI gene, we have demonstrated the existence of three different regions which are important for expression. One of these regions, which was localized to within 270 base pairs of the translation start codon, is involved in the periodic expression of TMPI transcript. A second region, the deletion of which resulted in reduced levels of TMPI expression, is at least partially encoded by DNA sequences between 270 and 377 base pairs upstream of the translation start codon. A third region, located within the N-terminal 112 codons of the TMPI gene, apparently encodes information involved in a post-translational control mechanism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419994
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  • 7
    Electronic Resource
    Electronic Resource
    Induced recombination and reversion of theCDC8 gene in relation to the cell cycle of yeast (1988)
    Springer
    Current genetics 13 (1988), S. 455-460 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Gene conversion and mutation ; CDC8 locus ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The induction of mitotic recombination in theCDC8 locus was studied in a diploid strain heteroallelic forcdc8 mutations (cdc8-1/cdc8-3); mitotic reversion was studied in strainscdc8-1/cdc8-1 andcdc8-3/cdc8-3. Conversion and reversion did not occur in those cells blocked at the S stage of the cell cycle by exposure to a nonpermissive temperature. In stationary phase cells irradiated just prior to exposure to temperature stress, the induction of recombinants was rather low and the induction of revertants was minimal. Conversely, a significant induction ofcdc + occurred in logarithmic phase cells subjected to the same treatment. Irradiation of synchronously dividing cultures revealed that intragenic recombination occurs at all three stages of the cell cycle- G1, S and G2. It was also found that UV-induced gene reversion can occur during the S and G2 stages, but not during the G1 stage of the cell cycle.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02427750
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  • 8
    Electronic Resource
    Electronic Resource
    A reexamination of the role of the RAD52 gene in spontaneous mitotic recombination (1988)
    Springer
    Current genetics 14 (1988), S. 211-223 
    Details
    ISSN: 1432-0983
    Keywords: Mitotic recombination ; DNA repair ; Yeast ; RAD52
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The RAD52 gene is required for much of the recombination that occurs in Saccharomyces cerevisiae. One of the two commonly utilized mutant alleles, rad52-2, increases rather than reduces mitotic recombination, yet in other respects appears to be a typical rad52 mutant allele. This raises the question as to whether RAD52 is really necessary for mitotic recombination. Analysis of a deletion/insertion allele created in vitro indicates that the null mutant phenotype is indeed a deficiency in mitotic recombination, especially in gene conversion. The data also indicate that RAD52 is required for crossing-over between at least some chromosomes. Finally, examination of the behavior of a replicating plasmid in rad52-1 strains indicates that the frequency of plasmid integration is substantially reduced from that in wild type, a conclusion consistent with a role for RAD52 in reciprocal crossing-over. Analysis of recombinants arising in rad52-2 strains suggests that this allele may result in the increased activity of a RAD52-independent recombinational pathway.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00376741
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  • 9
    Electronic Resource
    Electronic Resource
    Modifiers of ochre suppressors in Saccharomyces cerevisiae that exhibit ochre suppressor-dependent amber suppression (1988)
    Springer
    Current genetics 14 (1988), S. 345-354 
    Details
    ISSN: 1432-0983
    Keywords: Informational suppressors ; Modifier ; Yeast ; tRNAs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants of Saccharomyces cerevisiae were selected that would interact with ochre (UAA) suppressors so as to allow ochre -suppressor dependant amber (UAG) suppression, but which do not exhibit opal (UGA) suppression. Strains mutant at four distinct loci were isolated, and two of these are recessive mutations while the other two behave as dominants or semidominants. MOS3 has some suppressor activity in the absence of a resident SUP4-o gene and shares other characteristics with previously described omnipotent suppressors. MOS4, mos1 and mos2, on the other hand, exhibit no suppressor activity in the absence of a resident SUP4-o gene but do exhibit suppression of UAG alleles when there is a resident SUP4-o gene. These latter modifier strains do not interact with a SUP4-o gene to suppress UGA alleles. By genetic and physiological criteria the MOS4, mosl, and most mutations appear to be different than previously described allosuppressors or modifiers of suppression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419992
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  • 10
    Electronic Resource
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    Evidence that the single CDC8 gene which encodes thymidylate kinase is involved in DNA replication, recombination and mutation in yeast (1988)
    Springer
    Current genetics 14 (1988), S. 303-309 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; CDC8 gene ; DNA replication, recombination, mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The conditional cdc8 mutant is known to be defective, under restrictive conditions, in the elongation of DNA during synthesis. In yeast the CDC8 gene encodes thymidylate kinase. We show here that UV-induced gene conversion and gene mutation events require the participation of this CDC8 gene. Thus, the same thymidylate kinase is incolved both in DNA replication and in UV-induced gene conversion and gene mutation in yeast.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419986
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  • 11
    Electronic Resource
    Electronic Resource
    Carbon assimilation tests: Substrates assimilation profiles (1988)
    Springer
    Mycopathologia 102 (1988), S. 3-8 
    Details
    ISSN: 1573-0832
    Keywords: Yeast ; carbon assimilation profiles ; liquid medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Liquid medium assays for yeasts carbon assimilation tests are the more precise but longer methods. For rapid and automated yeasts identification purposes we analysed the assimilation of 34 carbon compounds by 149 reference strains. Assays were carried in liquid shaken medium (Autobac∘ system) and readings were nephelemetric. Valuable results are obtained in 72 hours and their analysis allowed us to classify substrates for their ability to minimize the number of doubtful results.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00436244
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  • 12
    Electronic Resource
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    Allelism between pso1-1 and rev3-1 mutants and between pso2-1 and snm1 mutants in Saccharomyces cerevisiaee (1988)
    Springer
    Current genetics 13 (1988), S. 37-40 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; DNA repair mutants ; Allelism test ; Psoralen plus UVA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the yeast Saccharomyces cerevisiae, allelism between the psol-1 and the rev3-1 mutants on the one hand and the pso2-1 and snm1 mutants on the other, is demonstrated by the comparison of phenotypes, complementation tests and meiotic segregation analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365754
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  • 13
    Electronic Resource
    Electronic Resource
    Interspecific protoplast fusion between the yeasts Saccharomyces cerevisiae and Saccharomyces fermentati (1988)
    Springer
    Current genetics 13 (1988), S. 101-104 
    Details
    ISSN: 1432-0983
    Keywords: Fusion ; Protoplast ; Saccharomyces ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts of Saccharomyces cerevisiae his1 trp2 resistant to acriflavine and able to ferment galactose and of Saccharomyces fennentati arg resistant to DL-p-fluorophenylalanine and able to ferment lactose were fused. As a result of fusion two types of prototrophic hybrids were obtained. Type 1 hybrids were able to grow on medium with galactose or lactose as sole carbon source and were sensitive to acriflavine and resistant to DL-p-pfluorophenylalanine. Type 2 hybrids were able to grow on medium with galactose as sole carbon source and were resistant to acriflavine and sensitive to DL-p-fluorophenylalanine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365764
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  • 14
    Electronic Resource
    Electronic Resource
    FLP recombinase induction of the breakage-fusion-bridge cycle and gene conversion in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 273-281 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; FLP-FRT ; BFBC ; Gene conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A YEp chimaeric plasmid containing URA3 and SMR1 [sulfometuron methyl resistant (SMR) allele of ILV2] as selectable markers, and the 2 μm site-specific recombination FLP recognition target (FRT), was integrated at the ilv2-Δ1 site in chromosome XIII in a cir°] haploid. Southern analysis defined two integrant structures. Structure I had URA3 distal and SMR1 proximal to FRT whereas in structure II both markers were distal to FRT. Selectable markers were stably inherited in [cir°] haploids and [cir°] diploids heterozygous for the integrant and ILV2. Approximately 14% of heterozygous [cir +] diploid cells exhibited homozygotization for the distal (500 kb) ade4 marker in trans. In [cir +] diploids FLP-FRT recombination resulted in the simultaneous loss of both structure II markers, whereas the structure I distal URA3 marker loss always preceded the variable loss of the proximal SMR1 marker. URA− cells continued to segregate for loss of SMR1 until stable URA− SMR or URA−SMS cells were produced. Gene conversion was identified in stable URA−SMR cells that were homozygous SMR1/SMR1 but contained wild type ILV2 restriction endonuclease sites. These observations support a model based on concerted FLP-FRT action resulting from the secondary integration of native 2 μm DNA followed by unequal sister chromatid exchange (USCE) within inverted FRTs. The resultant chromatid bridge resulted in a double-stand break. Fusion of the broken ends of sister chromatids generated a breakage-fusion-bridge cycle (BFBC). Repeated rounds of the BFBC resulted in proximal marker loss and the generation of additional double-strand breaks. Recombinogenic properties of the double-strand break initiated events leading to homozygotization and gene conversion.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00424420
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  • 15
    Electronic Resource
    Electronic Resource
    Sensitivity of yeast glycolytic enzymes to chloroquine (1988)
    Springer
    Archives of microbiology 150 (1988), S. 309-312 
    Details
    ISSN: 1432-072X
    Keywords: Chloroquine ; Glycolytic enzymes ; Yeast ; Chloroquine and ATP/ADP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chloroquine at pH 8.0 and 10 mM concentration inhibits about 30% glucose consumption and ethanol formation in yeast cells. Out of the 11 glycolytic enzymes assayed, phosphoglycerate kinase and pyruvate decarboxylase have been found to be most sensitive to chloroquine. Next sensitive are hexokinase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase. Kinetic studies with the three kinases studied revealed competitive inhibition of chloroquine with ATP (hexokinase, phosphoglycerate kinase) or ADP (pyruvate kinase).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00407797
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  • 16
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    Chloroquine, a lysosomotropic agent, inhibits zygote formation in yeast (1988)
    Springer
    Archives of microbiology 151 (1988), S. 20-25 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating ; Zygote formation ; Chloroquine ; Lysosomotropic agent ; Plasma membrane ; Cell fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Haploid cells of opposite mating type of Saccharomyces cerevisiae conjugate to form zygote. During the conjugation process, the degradation or reorganization of the cell wall and the fusion of the two plasma membranes take place. Since chloroquine inhibits cellular events associated with the reorganization of the plasma membrane, the effect of the drug on conjugation was studied. Chloroquine at a concentration, at which cell growth was not retarded, inhibited zygote formation, while it did not affect other mating functions, such as sexual agglutination, production of and response to mating pheromone. Cells in a mating culture containing chloroquine formed no “prezygote” suggesting that they were not prepared for entering into fusion process. The inhibitory effect of chloroquine was reversible as cells formed zygote when they were washed after treatment with chloroquine. Zygote formation was unaffected in cells possessing chlorquine within vacuoles after incubation with the drug in complete medium (YPD) at pH 7.5, followed by washing. This suggests that chloroquine inhibits zygote formaton by adsorbing to the plasma membrane of S. cerevisiae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00444663
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  • 17
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    How hexoses and inhibitors influence the malate transport system in Zygosaccharomyces bailii (1988)
    Springer
    Archives of microbiology 150 (1988), S. 37-41 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Hexose transport ; Sugar ; Malate uptake ; 2,4-DNP ; Zygosaccharomyces bailii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When grown in fructose or glucose the cells of Zygosaccharomyces bailii were physiologically different. Only the glucose grown cells (glucose cells) possessed an additional transport system for glucose and malate. Experiments with transport mutants had lead to the assumption that malate and glucose were transported by one carrier, but further experiments proved the existence of two separate carrier systems. Glucose was taken up by carriers with high and low affinity. Malate was only transported by an uptake system and it was not liberated by starved malate-loaded cells, probably due to the low affinity of the intracellular anion to the carrier. The uptake of malate was inhibited by fructose, glucose, mannose, and 2-DOG but not by non metabolisable analogues of glucose. The interference of malate transport by glucose, mannose or 2-DOG was prevented by 2,4-dinitrophenol, probably by inhibiting the sugar phosphorylation by hexokinase. Preincubation of glucose-cells with metabolisable hexoses promoted the subsequent malate transport in a sugar free environment. Preincubation of glucose-cells with 2-DOG, but not with 2-DOG/2,4-DNP, decreased the subsequent malate transport. The existence of two separate transport systems for glucose and malate was demonstrated with specific inhibitors: malate transport was inhibited by sodium fluoride and glucose transport by uranylnitrate. A model has been discussed that might explain the interference of hexoses with malate uptake in Z. bailii.
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    URL: http://dx.doi.org/10.1007/BF00409715
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  • 18
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    Arachidonic and oleic acids stimulate zygote formation in the presence of mating pheromone in the yeast, Saccharomyces cerevisiae (1988)
    Springer
    Archives of microbiology 149 (1988), S. 507-508 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating reaction ; Zygote formation ; Mating pheromone ; Fatty acid ; Arachidonic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effect of exogenous fatty acids on zygote formation in Saccharomyces cerevisiae was studied. Arachidonic and oleic acids considerably stimulated zygote formation, but other fatty acids tested, linoleic, linolenic, stearic and palmitic acids, did not. Pretreatment experiments with arachidonic acid showed that the stimulation of zygote formation by the fatty acid required the presence of mating pheromone.
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    URL: http://dx.doi.org/10.1007/BF00446752
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  • 19
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    Killer toxin producing strains of the yeasts Hanseniaspora uvarum and Pichia kluyveri (1988)
    Springer
    Archives of microbiology 149 (1988), S. 261-267 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Hanseniaspora uvarum ; Pichia kluyveri ; Killer toxin ; dsRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By heat treatment killer strains of the type K1 of Saccharomyces cerevisiae that are known to harbour dsRNA plasmids were completely cured, whereas only a small fraction of the clones of the killer type K2 had lost the dsRNA dependent killer character. The K2 killers but not the strains of killer type K1 were easily cured by cycloheximide. Killer strains of Hanseniaspora uvarum were not curable by heat treatment. Curing was successfull with cycloheximide or 5-fluorouracil. Two double-stranded RNA plasmids were detected in the killer strains of H. uvarum. The smaller dsRNA plasmid was absent in the strains that were cured of their killer character by 5-fluorouracil. The killer character of H. uvarum was transferred to S. cerevisiae by spheroplast fusion. The fusion products showing the killer character contained both dsRNA plasmids, obviously the smaller plasmid (M-dsRNA) carries the genes for killer toxin formation. Killer strains of Pichia kluyveri were not curable of their killer character, in these strains no dsRNA plasmids were detected.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422015
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  • 20
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    Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation (1988)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
    Details
    ISSN: 1476-5535
    Keywords: Osmotic pressure ; Intracellular ethanol ; Yeast ; Nutrient ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The intracellular accumulation of ethanol in yeast and its potential effects on growth and fermentation have been topics of controversy for the past several years. The determination of intracellular ethanol based on the exclusion of [14C]sorbitol to estimate aqueous cell volume was used to examine the question of intracellular ethanol accumulation. An intracellular accumulation of ethanol inSaccharomyces cerevisiae was observed during the early stages of fermentation. However, as fermentation continued, the intracellular and extracellular concentrations of ethanol became similar. Increasing the osmotic pressure of the medium with glucose or sorbitol was observed to cause an increase in the intracellular ethanol concentration. Associated with this was a decrease in yeast growth and fermentation rates. In addition, increasing the osmotic pressure of the medium was observed to cause an increase in glycerol production. Supplementation of the media with excess peptone, yeast extract, magnesium sulfate and potassium phosphate was found to relieve the detrimental effects of high osmotic pressure. Under these conditions, though, no effect on the intracellular and extracellular ethanol distribution was observed. These results indicate that nutrient limitation, and not necessarily intracellular ethanol accumulation, plays a key role during yeast fermentations in media of high osmolarity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569575
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  • 21
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    Induction of homologous recombination in Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 37-41 
    Details
    ISSN: 1617-4623
    Keywords: Homologous recombination ; UV induction ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have investigated the effects of UV irradiation of Saccharomyces cerevisiae in order to distinguish whether UV-induced recombination results from the induction of enzymes required for homologous recombination, of the production of substrate sites for recombination containing regions of DNA damage. We utilized split-dose experiments to investigate the induction of proteins required for survival, gene conversion, and mutation in a diploid strain of S. cerevisiae. We demonstrate that inducing doses of UV irradiation followed by a 6 h period of incubation render the cells resistant to challenge doses of UV irradiation. The effects of inducing and challenge doses of UV irradiation upon interchromosomal gene conversion and mutation are strictly additive. Using the yeast URA3 gene cloned in non-replicating single- and double-stranded plasmid vectors that integrate into chromosomal genes upon transformation, we show that UV irradiation of haploid yeast cells and homologous plasmid DNA sequences each stimulate homologous recombination approximately two-fold, and that these effects are additive. Non-specific DNA damage has little effect on the stimulation of, homologous recombination, as shown by studies in which UV-irradiated heterologous DNA was included in transformation/recombination experiments. We further demonstrate that the effect of competing single- and double-stranded heterologous DNA sequences differs in UV-irradiated and unirradiated cells, suggesting an induction of recombinational machinery in UV-irradiated S. cerevisiae cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340176
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  • 22
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    Precise excision of bacterial transposon Tn 5 in yeast (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 388-393 
    Details
    ISSN: 1617-4623
    Keywords: Tn5 ; Transposon excision ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have demonstrated that precise excision of bacterial transposon Tn5 can occur in the yeast, Saccharomyces cerevisiae. Tn5 insertions in the yeast gene LYS2 were generated by transposon mutagenesis made in Escherichia coli by means of a λ::Tn5 vector. Nine insertions of Tn5 into the structural part of the yeast LYS2 gene situated in a shuttle epsiomal plasmid were selected. All the plasmids with a Tn5 insertion were used to transform yeast strains carrying a deletion of the entire LYS2 gene or a deletion of the part of LYS2 overlapping the point of insertion. All insertions inactivated the LYS2 gene and were able to revert with low (about 10-8) frequencies to lysine prototrophy. Restriction analysis of revertant plasmids revealed them to be indistinguishable from the original plasmid without Tn5 insertion. DNA sequencing of the regions containing the points of insertions, made for two revertants, proved that Tn5 excision was completely precise.
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    URL: http://dx.doi.org/10.1007/BF00339607
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    The detection of mitotic and meiotic aneuploidy in yeast using a gene dosage selection system (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 10-18 
    Details
    ISSN: 1617-4623
    Keywords: Aneuploidy ; Yeast ; Saccharomyces cerevisiae ; Methyl benzimidazol-2-yl carbamate ; Mitosis ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A system is described in which spontaneous and chemically-induced mitotic and meiotic hyperploidy can be assayed in the same diploid culture of Saccharomyces cerevisiae. Monitoring gene dosage changes at two loci on chromosome VIII, the test utilizes a leaky temperature-sensitive allele arg4-8 and low level copper resistance conferred by the single copy allele cup1 s. An extra chromosome VIII provides simultaneous increased dosage for both genes, resulting in colonies that are both prototrophic for arginine at 30° C and copper resistant. During mitotic cell divisions in diploids, spontaneous chromosome VIII hyperploids (trisomes and tetrasomes) occur at a frequency of 6.4×10-6 per viable cell. Among ascospores, the spontaneous chromosome VIII disome frequency is 5.5×10-6 per viable spore. The tubulin-binding reagent methyl benzimidazol-2-yl carbamate (MBC) elicits enhanced levels of mitotic and meiotic aneuploidy relative to control levels. The system represents a novel model for examining chromosome behavior during mitosis and meiosis and provides a sensitive and quantifiable procedure for examining chemically induced aneuploidy.
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    URL: http://dx.doi.org/10.1007/BF00331296
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    Genetic characterization of hyperresistance to formaldehyde and 4-nitroquinoline-N-oxide in the yeast Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 260-265 
    Details
    ISSN: 1617-4623
    Keywords: Mutagen resistance ; Yeast ; Formaldehyde ; 4-Nitroquinoline-N-oxide ; Multi-copy plasmids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hyperresistance to 4-nitroquinoline-N-oxide (4-NQO) and formaldehyde (FA) of yeast strains transformed with the multi-copy plasmids pAR172 and pAR184, respectively, is due to the two genes, SNQ and SFA, which are present on these plasmids. Restriction analysis revealed the maximal size of SFA as 2.7 kb and of SNQ as 2.2 kb, including transcription control elements. The presence of the smallest 2.7 kb subclone carrying SFA increased hyperresistance to formaldehyde fivefold over that of the original pAR184 isolate. No such increase in hyperresistance to 4-NQO was seen with the smaller subclones of the pAR172 isolate. Disruption of the SFA gene led to a threefold increase in sensitivity to FA as compared with the wild type. Expression of gene SNQ introduced on a multi-copy vector into haploid yeast mutants rad2, rad3, and snm1 did not complement these mutations that block excision repair.
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    URL: http://dx.doi.org/10.1007/BF00330602
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    An endo-exonuclease activity of yeast that requires a functional RAD52 gene (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 41-48 
    Details
    ISSN: 1617-4623
    Keywords: RAD52 ; Repair ; Nuclease ; Antibody ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Extracts of Rad+ and radiation-sensitive (rad) mutants of the yeast Saccharomyces cerevisiae were examined for total Mg2+-dependent alkaline deoxyribonuclease activity and the presence of a nuclease that crossreacts immunologically with an antiserum raised against an endoexonuclease from Neurospora crassa, an enzyme exhibiting both deoxyribo- and ribonuclease activities. No significant differences were observed in total deoxyribonuclease activity between Rad+ and rad mutants. The antibody precipitable activity, however, was found to be 30%–40% of the total alkaline deoxyribonuclease activity in logarithmically growing Rad+ cells. Extracts of stationary phase cells were lacking in antibody precipitable activity. Using immunoblot methods, a 72 kDa crossreacting protein was identified from logarithmically growing cells that was absent from stationary phase cells. In all radiation-sensitive mutants examined, except rad52, at least 20% of total activity was precipitable. Extracts from logarithmically growing rad52 mutants, including a rad52::LEU2 insertion mutant, exhibited less than 10% of the Rad+ precipitable activity; however, some crossreacting material was detected. Although, the level of endo-exonuclease activity is influenced by the RAD52 gene, it is not the product of this gene. The total deoxyribonuclease and the antibody precipitable endo-exonuclease activities were also followed during meiosis. Unlike the Rad+ strain which had previously been shown to have increased levels of total and immunoprecipitable endo-exonuclease as cells underwent meiosis, the rad52 mutant exhibited no increases in either category of nuclease activity. Given the importance of the RAD52 gene in repair, recombination and mutagenesis, the endo-exonuclease may be a significant component of these processes.
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    URL: http://dx.doi.org/10.1007/BF00338391
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    Mutations in the NAM2 genes of Saccharomyces cerevisiae and S. douglasii are clustered non-randomly as a result of constraints on the nucleic acid sequence and not on the protein (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 310-314 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; tRNA synthetase gene ; Distribution of mutations ; Genetic drift
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present a statistical study of the nature and distribution of mutations along the NAM2 gene coding for the mitochondrial leucyl tRNA synthetase in Saccharomyces cerevisiae and S. douglasii (Herbert et al. 1988). Two important facts are observed: (1) the relative frequency of transitions and transversions is the same among silent substitutions and replacements. (2) The two kinds of mutations (silent substitutions and replacements) are distributed in the same way along the gene. This distribution is not random; the mutations are clustered and the clusters are regularly spaced along the gene. The NAM2 gene offers an example spaced along the gene. The NAM2 gene offers an example of recent divergence. We show that, in this case, the fixation of mutations is the result of genetic drift and of constraints on the nucleic acid sequence and not on that of the protein.
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    URL: http://dx.doi.org/10.1007/BF00339596
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    Divergence of the mitochondrial leucyl tRNA synthetase genes in two closely related yeasts Saccharomyces cerevisiae and Saccharomyces douglasii: A paradigm of incipient evolution (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 297-309 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; pre-mRNA splicing ; tRNA synthetase gene ; Incipient evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We studied the NAM2 genes of Saccharomyces douglasii and Saccharomyces cerevisiae, and showed that they are interchangeable for all the known functions of these genes, both mitochondrial protein synthesis and mitochondrial mRNA splicing. This confirms the prediction that the S. douglasii NAM2D gene encodes the mitochondrial leucyl tRNA synthetase (EC 6.1.1.4). The observation that these enzymes are interchangeable for their mRNA splicing functions, even though there are significant differences in the intron/exon structure of their mitochondrial genome, suggests that they may have a general role in yeast mitochondrial RNA splicing. A short open reading frame (ORF) precedes the synthetase-encoding ORF, and we showed that at least in S. cerevisiae this is not essential for the expression of the gene; however, it may be involved in a more subtle type of regulation. Sequence comparisons of S. douglasii and S. cerevisiae revealed a particularly interesting situation from the evolutionary point of view. It appears that the two yeasts have diverged relatively recently: there is remarkable nucleotide sequence conservation, with no deletions or insertions, but numerous (albeit non-saturating) silent substitutions resulting from transitions. This applies not only to the NAM2 coding regions, but also to two other ORFs flanking the NAM2 ORF. The regions between the ORFs (believed to be intergenic regions) are much less conserved, with several deletions and insertions. Thus S. douglasii and S. cerevisiae provide an ideal system for the study of molecular evolution, being two yeasts “caught in the act” of speciation.
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    URL: http://dx.doi.org/10.1007/BF00339595
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    Rad3 protein of Saccharomyces cerevisiae: Overexpression and preliminary characterization using specific antibodies (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 400-408 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; DNA repair ; RAD3 gene expression ; Fusion proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cloned RAD3 gene of Saccharomyces cerevisiae was tailored into expression vectors for overexpression of Rad3 protein in Escherichia coli and in yeast. In both organisms the overexpressed protein is detected as a species of molecular weight ca. 90 kDa, the size expected from the sequence of the cloned gene. The protein overexpressed in E. coli is largely insoluble; however the insoluble fraction was used to generate affinity-purified polyclonal antisera which proved to be powerful reagents for the initial characterization of Rad3 protein expressed in yeast. These studies showed that: (1) when overexpressed in yeast most of the Rad3 protein is detected in the soluble fraction of cell extracts; (2) endogenous Rad3 protein is untransformed cells is also ca. 90 kDa in size and is located in the cell nucleus; (3) Rad3/β-galactosidase fusion protein partially purified on an affinity matrix is associated with DNA-dependent ATPase activity that is inhibited in the presence of anti-Rad3 antibodies, suggesting that Rad3 protein is an ATPase; and (4) Rad3 antibodies cross-react with two electrophoretically distinguishable polypeptides present in the nuclear fraction of human cells, and with a single polypeptide in extracts of Drosophila cell.
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    URL: http://dx.doi.org/10.1007/BF00339609
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    Amplification of plasmid copy number by thymidine kinase expression in Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 155-159 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Yeast ; Copy number ; Thymidine kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2 μm circle-based chimaeric plasmid containing the yeast LEU2 and the Herpes Simplex Virus type 1 thymidine kinase (HSV-1 TK) genes was constructed. Transformants grown under selective conditions for the LEU2 gene harboured the plasmid at about 15 copies per cell whilst selection for the HSV-1 TK gene led to an increase to about 100 copies per cell. Furthermore, the plasmid copy number could be controlled by the stringency of selection for the TK gene, and the increase in TK gene dosage was reflected in an increase in intracellular thymidine kinase activity. The mitotic stability of the plasmid in “high-copy” and “low-copy” number cells was determined. “High-copy” number cells showed a greater mitotic stability. The relationship of TK expression to plasmid copy number may be useful for the isolation of plasmid copy number mutants in yeast and the control of heterologous gene expression.
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    URL: http://dx.doi.org/10.1007/BF00338407
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    The same configuration of Ty elements promotes different types and frequencies of rearrangements in different yeast strains (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 272-281 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Ty elements ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We examined Ty-mediated genomic rearrangements in three related mitotically dividing haploid yeast strains having the same configuration of Ty elements in the CYC1-sup4 interval of chromosome X. Surprisingly, quite different types and frequencies of rearrangements were found in the three strains. In one strain we found only Ty-mediated deletions, which occurred with a frequency of about 1×10-6. Another strain yielded similar deletions, but approximately one-third of these were accompanied by adjacent Ty-mediated inversions. A third strain was found to have an extremely high rate of inversion/reinversion between two of the three Ty elements. This rate was conservatively estimated to be 1.4±0.2×10-2 per cell per generation, which is at least 2 orders of magnitude higher than previously reported values for Ty-mediated rearrangements. These data provide evidence that local regions of the genome can, in some cases, be much more fluid than had been previously believed.
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    URL: http://dx.doi.org/10.1007/BF00330604
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    SCO1, a yeast nuclear gene essential for accumulation of mitochondrial cytochrome c oxidase subunit II (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 492-498 
    Details
    ISSN: 1617-4623
    Keywords: Cytochrome c oxidase ; Mitochondria ; PET genes ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified and isolated a novel yeast nuclear gene (SCO1) which is essential for accumulation of the mitochondrially synthesized subunit II of cytochrome c oxidase (CoxII). Analysis of the mitochondrial translation products in a sco1-1 mutant reveals a strong reduction in CoxII. Examination of mitochondrial transcripts by Northern blot hybridization shows that transcription and transcript maturation of OXI1, the gene coding for CoxII, is not affected. Therefore the SCO1 gene product must be involved in a post-transcriptional step in the synthesis of CoxII. We have isolated a 1.7 kb DNA fragment from a yeast gene bank which carries the functional SCO1 gene. Two RNA species of 0.9 kb and 1.2 kb, respectively, hybridize with this DNA fragment, which is localized on chromosome II. Cells whose chromosomal 1.7 kb fragment has been replaced by the yeast URA3 gene fail to accumulate CoxII and in addition subunit I of cytochrome c oxidase (CoxI). The possibility that the SCO1 gene product is bifunctional, i.e. required for both CoxI and CoxII accumulation, is discussed.
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    URL: http://dx.doi.org/10.1007/BF00425706
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    Genetic mapping of the Saccharomyces cerevisiae DNA polymerase I gene and characterization of a pol1 temperaturesensitive mutant altered in DNA primase-polymerase complex stability (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 459-465 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; DNA primase-polymerase complex ; Temperature sensitive mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cloned DNA polymerase I gene has been used to map the POL1 locus on the left arm of chromosome XIV, between MET4 and TOP2. Temperature-sensitive mutants in POL1 have been obtained by in vitro mutagenesis of the cloned gene and in vivo replacement of the wild-type allele with the mutated copy. Physiological and biochemical characterization of one temperature-sensitive mutant (pol1-1) shows that cells shifted to the non-permissive temperature can complete one round of cell division and DNA replication before they arrest. Analysis of DNA polymerase I in crude extracts and in partially purified preparations indicates that the pol1-1 mutation results in a conformational change and affects the stability of the DNA primase-polymerase complex.
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    URL: http://dx.doi.org/10.1007/BF00330850
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    Palindrome-hairpin linear plasmids possessing only a part of the ORF1 gene of the yeast killer plasmid pGKL1 (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 46-52 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; pGKL plasmids ; Palindrome-hairpin linear plasmids ; the ORF1 gene ; DNA polymerase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast Kluyveromyces lactis haboring linear DNA plasmids pGKL1 and pGKL2 exhibits killer and killer-resistant phenotypes. Two new linear plasmids pK192L and pK192S were found in the weak killer mutant KUV192 induced by UV irradiation. pK192S was always accompanied by pK192L in subclones of KUV192. Both plasmids were derived from pGKL1 by deletion of the large right part of it. pK192L was 4.9 kb in size and had a palindromic structure consisting of 2.35 kb inverted terminal repetitions and a 215 base unique sequence. Analysis of denatured and renatured DNA strands suggested that pK192S was a hairpin-like form of pK192L. The pK192 plasmids were maintained only in cells haboring either pGKL1 or pGKL1S in addition to pGKL2 and competed with pGKL1 or pGKL1S for their maintenance. Since no complete ORF1 was conserved in pK192 plasmids, these results lead to the conclusion that the ORF1 gene is necessary for the replication and/or maintenance of pGKL1.
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    URL: http://dx.doi.org/10.1007/BF00331301
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    The structure and regulation of phosphoglucose isomerase in Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 100-106 
    Details
    ISSN: 1617-4623
    Keywords: Phosphoglucose isomerease ; Regulation ; Metabolism ; Glycolysis ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned and sequenced the PGI1 gene, encoding phosphoglucose isomerase (E.C.5.3.1.9), from Saccharomyces cerevisiae. The nucleotide sequence predicts subunits of 554 amino acids with a molecular weight of 61230. Both the size and amino acid composition correlate well with measurements from purified protein. We have compared the PGI1 protein with the predicted sequence for pig muscle PGI. In spite of some evolutionary divergence the proteins are very similar and there are some highly conserved regions, two of which have been implicated in the active site. It has been suggested that PGI exists in two or more isozyme forms in S. cerevisiae and analogy with ADR2/ADC1 suggests that such PGI isozymes might also be differentially regulated during glycolytic/gluconeogenic growth. We have used accurate quantitation of PGI1 mRNA and gene fusions of PGI1 to the lacZ gene of Escherichia coli to show that PGI1 transcription is regulated neither between glycolytic and gluconeogenic growth nor between exponential and stationary phase. The complete lack of PGI activity in PGI1 deletion mutants and of differential regulation suggests that the isozymes of PGI might result merely from processing of the PGI1 gene product.
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    URL: http://dx.doi.org/10.1007/BF00331310
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