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  • Articles  (162)
  • Yeast  (162)
  • Springer  (162)
  • Institute of Physics
  • 1985-1989  (162)
  • 1950-1954
  • 1925-1929
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  • Articles  (162)
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  • 1
    Electronic Resource
    Electronic Resource
    Effects of nucleotides and divalent cations on phospholipase activity in Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 154-158 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Yeast ; Phospholipase B ; Lysophospholipase ; Enzyme inhibition ; AMP ; Unesterified fatty acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Divalent cations activate the lysophospholipase and transacylase reactions catalyzed by the same enzymes in the yeast Saccharomyces cerevisiae. The activation was observed at neutral pH, but not at the pH optimum of lysophospholipase/transacylase, near 3.5. Adenine nucleotides, especially AMP and ADP, are strong inhibitors of the same group of enzymes. Half maximal inhibition by AMP was found at a concentration of about 20 μM. The inhibition by nucleotides in low concentrations is enhanced by divalent cations.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00414431
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  • 2
    Electronic Resource
    Electronic Resource
    Genetic analysis of inducible sexual agglutination ability in the yeast Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 198-202 
    Details
    ISSN: 1432-072X
    Keywords: Sexual agglutination ; Mating ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic regulation of the inducibility of sexual agglutination ability in the yeast Saccharomyces cerevisiae was studied. Detailed analysis of the degree of sexual agglutination was carried out; it showed that a greater number of genes are involved in the regulation of inducible sexual agglutination in strain H1-0 than previously assumed. Although dominancy of inducible phenotype over constitutive was confirmed, the effectiveness of one gene changing the constitutive phenotype to the inducible seemed to be somewhat low. Quantity per cell of agglutination substances responsible for sexual agglutination increased as the agglutination ability became greater.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00413130
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  • 3
    Electronic Resource
    Electronic Resource
    A yeast ribosomal DNA-binding protein that binds to the rDNA enhancer and also close to the site of Pol I transcription initiation is not important for enhancer functioning (1989)
    Springer
    Current genetics 16 (1989), S. 351-359 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Transcription ; RNA polymerase I ; Enhancer ; DNA-binding protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using the gel retardation assay we have identified a protein that can specifically bind to a site within the enhancer of the 37S pre-ribosomal RNA operon in yeast, as well as to a site 210 by upstream of the site of transcription initiation of this operon. This protein (RBP1) has been partially purified by means of heparin-agarose chromatography and protects 20 by in the rDNA enhancer, and 25 by in the initiation region, against DNase I in an in vitro footprinting assay. In vivo footprinting studies using methylation of intact yeast cells with dimethylsulphate, indicate that the same binding sites are occupied in vivo as well. Deletions that abolish binding of RBP1 to the enhancer in vitro, as well as linker insertions into the RBP1 binding site in the initiation region that strongly diminish in vitro binding of RBP1, have no effect whatsoever on the enhancement of rDNA transcription in vivo. This was studied by deletion/mutation of the RBP1 binding site in vitro in an artificial ribosomal minigene and measuring the effect on the minigene transcription in vivo in yeast cells, transformed with the deleted/mutated minigenes. It can therefore be concluded that binding of RBP1 is not an important parameter in the functioning of the rDNA enhancer in yeast. Using the same minigene system we also show that RBP1 is not involved in termination of RNA polymerase I (PolI) transcription at the main terminator T2.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340714
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  • 4
    Electronic Resource
    Electronic Resource
    Two new loci that give rise to dominant omnipotent suppressors in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 323-330 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Nonsense suppression ; Omnipotent suppressors ; Gene mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ten dominant omnipotent suppressors of Saccharomyces cerevisiae, which were previously shown to be different from SUP46, have been examined. Nine are mapped in a region between lys5 and cyh2 on the left arm of chromosome VII. These suppressors, like SUP46, manifest sensitivity to increased temperature and the antibiotics paromomycin and hygromycin B. In addition, they have an identical action spectrum. These results strongly suggest that they are allelic to each other and they are designated SUP138. The tenth is mapped to a position between his1 and arg6 on the right arm of chromosome V. This suppressor, named SUP139, does not manifest temperature sensitivity nor antibiotic sensitivity. SUP139 and SUP138, which are clearly distinguished by means of action spectrum, act on much fewer nonsense mutations than SUP46. It is now clear that dominant omnipotent suppressors arising at a single locus are homogeneous and that their efficiency is locus-dependent. The order of efficiency is SUP46〉SUP138〉SUP139.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340710
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  • 5
    Electronic Resource
    Electronic Resource
    High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier (1989)
    Springer
    Current genetics 16 (1989), S. 339-346 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Transformation ; ss carrier DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A method, using LiAc to yield competent cells, is described that increased the efficiency of genetic transformation of intact cells of Saccharomyces cerevisiae to more than 1 × 105 transformants per microgram of vector DNA and to 1.5% transformants per viable cell. The use of single stranded, or heat denaturated double stranded, nucleic acids as carrier resulted in about a 100 fold higher frequency of transformation with plasmids containing the 2μm origin of replication. Single stranded DNA seems to be responsible for the effect since M13 single stranded DNA, as well as RNA, was effective. Boiled carrier DNA did not yield any increased transformation efficiency using spheroplast formation to induce DNA uptake, indicating a difference in the mechanism of transformation with the two methods.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340712
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  • 6
    Electronic Resource
    Electronic Resource
    Formation and stability of interstrand cross-links induced by cis- and trans-diamminedichloroplatinum (II) in the DNA of Saccharomyces cerevisiae strains differing in repair capacity (1989)
    Springer
    Current genetics 16 (1989), S. 331-338 
    Details
    ISSN: 1432-0983
    Keywords: Platinum compounds ; Yeast ; Repair mutants ; Interstrand cross-links ; DNA degradation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four haploid yeast strains differing in proficiency for DNA repair were treated with cis- or transDDP. The wild type was least sensitive while the excision-deficient mutants rad1, rad2 and snm1exhibited higher sensitivities to either platinum compound. In all four strains tested cisDDP showed a two- to five-fold higher cytotoxicity than equimolar concentrations of transDDP. DNA interstrand cross-linking was caused by both agents in all strains. However, transDDP introduced more DNA cross-links at exposure times up to 6 h while cisDDP was the more active cross-linking agent at longer times. There was no clear-cut correlation of the number of DNA interstrand cross-links with survival. Formaldehyde-treated cells showed DNA with lower buoyant density due to proteinase K sensitive DNA-protein cross-linking; this effect was not observed after treatment with either platinum compound. Post-treatment incubation of wild-type cells exposed to cisDDP led to degradation of DNA by single and double-strand breaks, parallel with further increase of DNA interstrand cross-linking. DNA from transDDP-treated cells did not show extensive degradation although interstrand cross-links were lost during liquid holding.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340711
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  • 7
    Electronic Resource
    Electronic Resource
    Two genes encode 7SL RNAs in the yeast Yarrowia lipolytica (1989)
    Springer
    Current genetics 16 (1989), S. 347-350 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; 7SL RNA ; Yarrowia lipolytica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified an abundant cytoplasmic 7S RNA in crude extracts of the yeast Yarrowia lipolytica. A cDNA probe was prepared from this RNA and used to screen a genomic library. The DNA sequence of a positive clone was determined and the end positions of the 7S RNA gene established by comparison with the sequence of the extremities of 7S RNA. This gene, designated SCR2, encodes a 270-nucleotide RNA that can be folded into a secondary structure similar to that of 7SL RNAs. This RNA is 94.4% homologous to a previously identified 7S RNA from this yeast, but is encoded by a separate gene with highly divergent flanking sequences.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340713
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  • 8
    Electronic Resource
    Electronic Resource
    Two nuclearly inherited loci conferring increased diuron resistance to NADH oxidase in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 31-38 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Diuron ; Respiration ; Nuclear genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, diuron blocks the respiration pathway at the level of the bc1 complex. Nuclear diuron-resistant mutations which confer in vitro resistance to mitochondrial NADH oxidase have been identified. Five mutations were found to be clustered at two distinct nuclear loci, DIU3 and DIU4. The distance between the two loci was estimated to be about 36.7 cM. These loci do not appear to be centromere-linked and did not show a linkage to any of the genes coding for bc1 complex subunits. DIU3 and DIU4 loci might, therefore, code for other components of the respiratory chain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00445749
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  • 9
    Electronic Resource
    Electronic Resource
    A deletion of the PDC1 gene for pyruvate decarboxylase of yeast causes a different phenotype than previously isolated point mutations (1989)
    Springer
    Current genetics 15 (1989), S. 75-81 
    Details
    ISSN: 1432-0983
    Keywords: Alcoholic fermentation ; Deletion mutant ; Pyruvate decarboxylase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We deleted most of the pyruvate decarboxylase structural gene PDC1 from the genome of Saccharomyces cerevisiae. Surprisingly, mutants carrying this deletion allele showed a completely different phenotype than previously described point mutations. They were able to ferment glucose and their specific pyruvate decarboxylase activity was only reduced to 45% of the wild type level. Northern blot analysis revealed that a sequence in the yeast genome homologous to PDC1 and formerly designated as a possible pseudogene is expressed and may code for a different but closely related pyruvate decarboxylase. The products of the two PDC genes seem to form hybrid oligomers, however both homooligomers have enzyme activity. Thus, the product of the PDC1 gene is not absolutely neccessary for glucose fermentation in yeast.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435452
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  • 10
    Electronic Resource
    Electronic Resource
    A novel class of vector for yeast transformation (1989)
    Springer
    Current genetics 16 (1989), S. 21-25 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Vectors ; Stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have constructed a set of hybrid yeast Escherichia coli vectors which utilise the site specific recombination function of the Saccharomyces cerevisiae 2 μm plasmid to completely eliminate the bacterial moiety upon introduction into yeast. A number of these plasmids have been shown to exhibit high inheritable stability in both laboratory and industrial strains during non-selective growth. These plasmids are beneficial for the genetic modification of industrial yeast, particularly those used in the production of food and beverages, and are of benefit in the study of plasmid maintenance and heterologous gene expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00411079
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  • 11
    Electronic Resource
    Electronic Resource
    Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: localization of a repeated sequence containing an acid phosphatase gene near a telomere of chromosome I and chromosome VIII (1989)
    Springer
    Current genetics 16 (1989), S. 131-137 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Chromosome organization ; Acid phosphatase ; Telomere
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 17 kb region from near the right end of chromosome I of Saccharomyces cerevisiae was isolated on recombinant λ bacteriophages. This region contained the PH011 gene which was located only 3.4 kb from the right end of the chromosome. We found that this region also was repeated approximately 13 kb from the end of the chromosome VIII DNA molecule. The chromosome VIII sequence appears to be a previously unnamed acid phosphatase gene that we propose to call PH012. Thus, similar to the repeated SUC, MAL, X and Y' sequences, some members of the repeated acid phosphatase gene family also appear near the termini of yeast chromosomes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00391468
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  • 12
    Electronic Resource
    Electronic Resource
    The UV excision-repair system of Saccharomyces cerevisiae is involved in the removal of methylcytosines formed in vivo by a cloned prokaryotic DNA methyltransferase (1989)
    Springer
    Current genetics 16 (1989), S. 461-464 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; DNA methylation ; DNA methyltransferase ; rad mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA methyltransferase activity is not normally found in yeast. To investigate the response of Saccharomyces cerevisiae to the presence of methylated bases, we introduced the Bacillus subtilis SPR phage DNA-[cytosine-5] methyltransferase gene on the shuttle vector, YEp51. The methyltransferase gene was functionally expressed in yeast under the control of the inducible yeast GAL10 promoter. Following induction we observed a time-dependent methylation of yeast DNA in RAD + and rad2 mutant strains; the rad2 mutant is defective in excision-repair of UV-induced DNA damage. Analysis of restriction endonuclease digestion patterns revealed that the relative amount of methylated DNA was greater in the excision defective rad2 mutant than in the RAD + strain. These data indicate that the yeast excision-repair system is capable of recognizing and removing m5C residues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340726
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  • 13
    Electronic Resource
    Electronic Resource
    Identification, cloning and characterisation of a new gene required for full pyruvate decarboxylase activity in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 171-175 
    Details
    ISSN: 1432-0983
    Keywords: PDC3 ; Pyruvate decarboxylase ; Subunits ; Yeast ; Cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Biochemical evidence that pyruvate decarboxylase in S. cerevisiae might be constituted from two independently encoded subunits led us to question genetic evidence for a single structural gene. The main evidence for this was that three “structural” mutations appeared to be alleles of the same gene, PDC1 (Schmitt and Zimmermann 1982). We report that one of these mutations (pdcl-30) is not allelic either to other pdc1 alleles or to pdc2 mutations and therefore is has been renamed pdc3-30 thus identifying a new gene, PDC3. We have cloned the PDC3 gene, it represents a unique sequence in the genome and targeted integration shows tight linkage to the PDC3 locus. However, the size, abundance and regulation of the PDC3 transcript suggest that it does not encode a second structural gene. Possible functions for the PDC3 gene product are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435502
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  • 14
    Electronic Resource
    Electronic Resource
    An α-specific gene, SAG1 is required for sexual agglutination in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 393-398 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mating ; Sexual agglutination ; a-Specific mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Seven α-specific mutants specifically defective in sexual agglutinability were isolated. The other α mating functions exhibited by these mutants, designated sag mutants, such as the production of α pheromone and response to a mating pheromone, were normal. While the MATα sag1 cells did not agglutinate with wild-type a cells, the MATα sag1 cells did, indicating that the SAG1 gene is expressed only in α cells. The mutations were semi-dominant and fell into a single complementation group, SAG1, which was mapped near met3 on chromosome X. Complementation analysis showed that sag1 and aga1, the latter being a previously reported α-specific mutation, were mutations in the same gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00376793
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  • 15
    Electronic Resource
    Electronic Resource
    Meiotic segregation of circular plasmid-minichromosomes from intact chromosomes in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 385-392 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Meiosis ; Distributive disjunction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Distributive disjunction is defined by first meiotic division segregation of either two nonhomologous chromosomes that lack homologous pairing partners, or of two homologous chromosomes that have failed to undergo crossing-over. In the yeast Saccharomyces cerevisiae, plasmid minichromosomes, synthetic linear chromosomes and a fragment of a real chromosome have been observed to segregate from nonhomologous DNA species at the first meiotic divisions. Suggesting that this organism may have a distributive mechanism for chromosome segregation. However, it is not known whether intact chromosomes also participate in a distributive process. To determine whether intact, full length, S. cerevisiae chromosomes could segregate from nonhomologous chromosomal species, the meiotic behavior of an unpaired intact copy of chromosome I has been analyzed with respect to several centromere-containing circular plasmid minichromosomes. Strains monosomic or trisomic for chromosome I were transformed with centromere plasmids containing either homologous or nonhomologous inserts, sporulated, and analyzed genetically both for the presence of plasmid and for the number of copies of chromosome 1. Each plasmid segregated from an intact unpaired copy of chromosome I at the first meiotic division in a significant majority (63–93%) of the asci examined. These results suggest that intact chromosomes from S. cerevisiae are capable of distributive disjunction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00376792
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  • 16
    Electronic Resource
    Electronic Resource
    A mitochondrial frameshift suppressor maps in the tRNASer-var1 region of the mitochondrial genome of the yeast S. cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 239-246 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial frameshift suppressor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A polypeptide chain-terminating mutation (M5631) previously has been shown to be a +1T insertion in the yeast mitochondrial gene oxi1, coding for subunit II of the cytochrome c oxidase. A spontaneously arisen frameshift suppressor (mfs-1) that is mitochondrially inherited suppresses this mutation to a considerable extent. The suppressor mutation was mapped by genetic and molecular analyses in the mitochondrial tRNASer-var1 region of the mitochondrial genome of the yeast S. cerevisiae. Genetic analyses show that the suppressor mfs-1 does not suppress other known mitochondrial frameshift mutations, or missense and nonsense mutations.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00447038
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  • 17
    Electronic Resource
    Electronic Resource
    Mutational analysis of the upstream activation site of yeast ribosomal protein genes (1989)
    Springer
    Current genetics 15 (1989), S. 247-251 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Ribosomal protein gene ; Transcription activation ; Mutation ; Methylation interference
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Most ribosomal protein (rp-)genes in yeast are preceded by conserved sequence motifs that act as upstream transcription-activating sites (RPG box). These sequence elements have previously been shown to represent specific binding sites for a protein factor, TUF. Comparison of the various nucleotide elements identified so far indicates a remarkably high degree of variation in the respective sequences. On the other hand, a methylation interference study performed with one RPG box revealed close contact points with the TUF protein along the entire sequence. To investigate the sequence requirements of the RPG box, we inserted synthetic oligonucleotides that differed from the general consensus sequence ACACCCATACATTT at single positions into a deletion mutant of the L25 promoter that lacked its natural RPG elements. Transcription activity was estimated by Northern analyses of the cellular level of L25-galK hybrid transcripts. The results show that in the 3′ part of this sequence element single substitutions are allowed at all positions, in the 5′ part, however, the nucleotide requirements appear to be more stringent. In particular, the invariant C at position 5 of the consensus sequence is absolutely necessary for its enhancer function.
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    URL: http://dx.doi.org/10.1007/BF00447039
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  • 18
    Electronic Resource
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    A controversy concerning the structure of the mitochondrial genome of a yeast petite mutant: resolution by sequence analysis (1989)
    Springer
    Current genetics 15 (1989), S. 291-293 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; oxi3 gene ; Petite genome ; Frameshift mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sequence analysis was used to define the repeat unit that constitutes the mitochondrial genome of a petite (rho −) mutant of the yeast Saccharomyces cerevisiae. This mutant has retained and amplified in tandem a 2,547 by segment encompassing the second exon of the oxi3 gene excised from wild-type mtDNA between two direct repeats of 11 nucleotides. The identity of the mtDNA segment retained in this petite has recently been questioned (van der Veen et al., 1988). The results presented here confirm the identity of this mtDNA segment to be that determined previously by restriction mapping (Carignani et al., 1983).
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    URL: http://dx.doi.org/10.1007/BF00447045
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  • 19
    Electronic Resource
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    Base substitutions in the 5′ non-coding regions of two naturally occurring yeast invertase structural SUC genes cause strong differences in specific invertase activities (1989)
    Springer
    Current genetics 15 (1989), S. 299-301 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Invertase ; Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Gene SUC4 produced about four fold more invertase activity than did gene SUC5. However, these genes differ in only three positions located in the 5′ non-coding region. The difference in gene expression between SUC4 and SUC5 must be due to the G to A transition (position −497) and/or the C to T transition (position −460) in the upstream activator sequences. The sequence TACAAA present in SUC5 can play the same role than the TATAAA box of SUC4.
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    URL: http://dx.doi.org/10.1007/BF00447048
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  • 20
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    The Saccharomyces cerevisiae RAD2 gene complements a Schizosaccharomyces pombe repair mutation (1989)
    Springer
    Current genetics 15 (1989), S. 27-30 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Repair ; Complementation ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two Saccharomyces cerevisiae genes necessary for excision repair of UV damage in DNA, RAD1 and RAD2, were introduced individually, on a yeast shuttle vector, into seven Schizosaccharomyces pombe mutants — rads1, 2, 5, 13, 15,16 and 17. The presence of the cloned RAD1 gene did not affect survival of any of the S. pombe mutants. The RAD2 gene increased survival of S. pombe rad13 to near the wild-type level after UV irradiation and had no effect on any of the other mutants tested. S. pombe rad13 mutants are somewhat defective in removal of pyrimidine dimers so complementation by the S. cerevisiae RAD2 gene suggests that the genes may code for equivalent proteins in the two yeasts.
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    URL: http://dx.doi.org/10.1007/BF00445748
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  • 21
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    Isolation and complete sequence of the yeast isoleucyl-tRNA synthetase gene (ILS1) (1989)
    Springer
    Current genetics 15 (1989), S. 99-106 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Isoleucyl-tRNA synthetase ; Isoleucine ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The isoleucyl-tRNA synthetase gene (ILS1) from the yeast Saccharomyces cerevisiae was cloned and sequenced. This gene was initially cloned because it cross-hybridizated to what is now presumed to be the isoleucyl-tRNA synthetase gene (cupC) from the protozoan Tetrahymena hhermophila. The ILS1 gene was determined to be 1,072 amino acids in length. A comparison with a recently published sequence of ILS1 1 from another laboratory (Englisch et al. 1987) was made and differences noted. Two promoter elements were detected, one for general amino acid control and one for constitutive transcription. A heat shock protein (hsp70) gene (probably SSA3) was found 237 by upstream from the ILS1 translation start site. The ILS1 amino acid sequence was compared to isoleucyl-tRNA synthetases from other organisms, as well as to valyl-, leucyl- and methionyl-tRNA synthetases. Regions of conservation between these enzymes were found.
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    URL: http://dx.doi.org/10.1007/BF00435455
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  • 22
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    A direct selection procedure for isolating yeast mutants with an impaired segregation of artificial minichromosomes (1989)
    Springer
    Current genetics 15 (1989), S. 17-25 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Minichromosomes ; Impaired segregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nondisjunction of artificial yeast minichromosomes (2:0 segregation events) during mitosis is accompanied by the appearance of cells containing more than one copy of the mini-chromosome. A mathematical simulation of this process has demonstrated that under certain conditions, a nondisjunction of the minichromosomes may result in their accumulation in a considerable portion of the cell population. An increase in the copy number of artificial minichromosomes as a result of impaired segregation has been used to develop a new experimental procedure for directly selecting yeast mutants showing an impaired segregation of artificial minichromosomes during mitosis. Four new genes, AMC1, AMC2, AMC3, and AMC4, which control the segregation of artificial minichromosomes in mitosis, have been identified (AMC-3 and AMC4 are mapped to chromosome IV and VII, respectively). Mutations in the genes AMC1–AMC4 also affect the mitotic transmission of natural chromosomes. We suggest that the genes AMC1, AMC2, AMC3, and AMC4 control the segregation of natural chromosomes in yeast.
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    URL: http://dx.doi.org/10.1007/BF00445747
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  • 23
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    FLP-FRT mediated intrachromosomal recombination on a tandemly duplicated YEp integrant at the ILV2 locus of chromosome XIII in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 107-112 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; 2μm FRT duplication ; Intrachromosomal recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A YEp chimaeric plasmid carrying SMR1 and URA3 genetic markers was integrated into chromosome XIII at the ilv2-Δ1 locus in a [cir°] background. The 1.5 kb BglII deletion of ilv2-Δ1 allowed the clear identification of an integrant structure which consisted of a direct tandem duplication (TD) of the chimaeric plasmid. Within the integrant structure, a single copy of the plasmid sequence was flanked by a direct duplication of the 2μm site-specific recombinase (FLP) recognition target (FRT). Isogenic [cir°] and [cir +] diploids formed by crossing the [cir°] TD strain to complementary haploids were analyzed for plasmid marker loss and chromosomal DNA alterations in the presence and absence of selection pressure for the URA3 and SMR1 plasmid borne markers. [cir°] diploids showed no plasmid marker loss and maintained the TD structure. In the absence of selection pressure, the [cir +] diploid underwent FLP-FRT mediated unequal interchromatid recombination, resulting in the breakage-fusion-bridge cycle and homozygotization of chromosome XIII (Rank et al. 1988). Maintenance of selection pressure for the centromere distal plasmid URA3 marker selected against FLP-FRT interchromatid recombinants so that the effects of site specific recombinase on intrachromatid recombination could be evaluated. Intrachromatid recombination at the directly duplicated FRT sites of the TD structure resulted in the loss of a diagnostic internal fragment. These results show that in the presence of FLP, FRT sites separated by up to 13.3 kb of chromosomal DNA function as substrates for intra and interchromatid recombination.
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    Increased diuron resistance in the joint expression of mutations located at the DIU2, DIU3 and DIU4 loci of Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 121-127 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Diuron ; Nuclear, mitochondrial mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, diuron blocks the respiratory pathway at the level of the bc1 complex. Two mitochondrially inherited loci, DIU1 and DIU2, located in the cytochrome b gene, and two nuclearly inherited loci, DIU3 and DIU4, have previously been identified. The present work genetically characterizes two double mutants. One mutant, Diu-217, carries two nuclearly inherited mutations, diu3-217a and diu-217b; the second mutant, Diu-783, carries the previously described nuclear mutation diu3-783 and a mitochondrial mutation diu2-783. Each mutation, independent of its location, exhibits a weak diuron resistance. The joint expression of two or three mutations leads to a cumulative or a cooperative enhanced diuron-resistant phenotype.
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    Synthesis and function of the mitochondrial intron —encoded bI4 RNA maturase from Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 241-246 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Intron splicing ; RNA maturase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed the expression and function of the intron-encoded bI4 maturase when frame-shift mutations in the upstream exon alter the translational process. By constructing secondary cis-acting mutations within the b14 intron, we observed (1) that the bI4 maturase is still translated in the presence of the upstream mutation, albeit in very low amounts, and (2) that the limited amounts of bI4 maturase made under these conditions is no longer able to promote the splicing process of the aI4 intron. These observations, which further strengthen the maturase model, strongly suggest that bI4 maturase acts sequentially on the bI4 intron and then on the aI4 intron.
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    URL: http://dx.doi.org/10.1007/BF00422109
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    A yeast Telomere Binding Activity binds to two related telomere sequence motifs and is indistinguishable from RAPT (1989)
    Springer
    Current genetics 16 (1989), S. 225-239 
    Details
    ISSN: 1432-0983
    Keywords: Telomere Binding Activity (TBA) ; Yeast ; Telomeric binding sites ; RAP1 gene product
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Telomere Binding Activity (TBA), an abundant protein from Saccharomyces cerevisiae, was identified by its ability to bind to telomeric poly(C1–3A) sequence motifs. The substrate specificity of TBA has been analyzed in order to determine whether the activity binds to a unique structure assumed by the irregularly repeating telomeric sequences or whether the activity recognizes and binds to subset of specific sequences found within the telomere repeat tracts. Deletion analysis and DNase I protection assays demonstrate that TBA binds specifically to two poly(C1–3A) sequences that differ by one nucleotide. The methylation of four guanine residues, located at identical relative positions within these two binding sequences, interferes with TBA binding to the substrates. A synthetic olignucleotide containing a single TBA binding site can function as a TBA binding substrate. The TBA binding site shares homology with the binding sites reported for the Repressor/Activator Protein 1 (RAP1), Translation Upshift Factor (TUF) and General Regulatory Factor (GRFI) transcription factors, and TBA binds directly to RAP1/TUF/GRFI substrate sequences. Yeast TBA preparations and the RAP1 gene product expressed in E. coli cells are both similarly sensitive to in vitro protease digestion. Affinity-purified TBA extracts include a protein indistinguishable from RAP1 in binding specificity, size, and antigenicity. The binding affinity of TBA for the two telomeric poly(C1–3A) binding sites is higher than its affinity for any of the other binding substrates used for its identification. In extracts of yeast spheroplasts prepared by incubation of yeast cells with Zymolyase, an altered, proteolyzed form, of TBA (TBA-S) is present. TBA-S has a faster mobility in gel retardation assays and SDS-PAGE gels, yet it retains the DNA binding properties of standard TBA preparations: it binds to RAP1/TUF/GRFI substrates with the same relative binding affinity and protects poly(C1–3A) tracts from DNase I digestion with a “footprint” identical to that of standard TBA preparations.
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    A malic acid dependent mutant of Schizosaccharomyces malidevorans (1989)
    Springer
    Archives of microbiology 152 (1989), S. 564-566 
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    ISSN: 1432-072X
    Keywords: l-Malate ; Schizosaccharomyces malidevorans ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The yeast Schizosaccharomyces malidevorans utilizes l-malate when grown on glucose as the carbon source. A mutant of this yeast has been isolated which is dependent on the presence of both l-malate and glucose for growth. The mutant utilizes l-malate as rapidly as the wildtype and the utilization of glucose is greatly reduced. Other TCA cycle intermediates do not relieve the malate dependence.
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    Effect of aerobiosis on fermentation and key enzyme levels during growth of Pichia stipitis, Candida shehatae and Candida tenuis on d-xylose (1989)
    Springer
    Archives of microbiology 152 (1989), S. 143-147 
    Details
    ISSN: 1432-072X
    Keywords: d-Xylose fermentation ; Aeration level ; Xylose reductase ; Xylitol dehydrogenase ; Yeast ; Candida shehatae ; Candida tenuis ; Pichia stipitis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between the degree of aerobiosis, xylitol production and the initial two key enzymes of d-xylose metabolism were investigated in the yeasts Pichia stipitis, Candida shehatae and C. tenuis. Anoxic conditions severely curtailed growth and retarded ethanol productivity. This, together with the inverse relationship between xylitol accumulation and aeration level, suggested a degree of redox imbalance. The ratios of NADH- to NADPH-linked xylose reductase were similar in all three yeasts and essentially independent of the degree of aerobiosis, and thus did not correlate with their differing capacities for ethanol production, xylitol accumulation or growth under the different conditions of aerobiosis. Under anoxic conditions the enzyme activity of Pichia stipitis decreased significantly, which possibly contributed to its weaker anoxic fermentation of xylose compared to C. shehatae.
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    Sugar utilization by yeast during fermentation (1989)
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    Journal of industrial microbiology and biotechnology 4 (1989), S. 315-323 
    Details
    ISSN: 1476-5535
    Keywords: Sugar uptake ; Yeast ; Brewer's wort
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary When glucose and fructose are fermented separately, the uptake profiles indicate that both sugars are utilized at similar rates. However, when fermentations are conducted in media containing an equal concentration of glucose and fructose, glucose is utilized at approximately twice the rate of fructose. The preferential uptake of glucose also occurred when sucrose, which was first rapidly hydrolyzed into glucose and fructose by the action of the enzyme invertase, was employed as a substrate. Similar results were observed in the fermentation of brewer's wort and wort containing 30% sucrose and 30% glucose as adjuncts. In addition, the high levels of glucose in the wort exerted severe catabolite repression on maltose utilization in theSaccharmyces uvarum (carlsbergensis) brewing strain. Kinetic analysis of glucose and fructose uptake inSaccharomyces cerevisiae revealed aK m of 1.6 mM for glucose and 20 mM for fructose. Thus, the yeast strain has a higher affinity for glucose than fructose. Growth on glucose or fructose had no repressible effect on the uptake of either sugar. In addition, glucose inhibited fructose uptake by 60% and likewise fructose inhibited, glucose uptake by 40%. These results indicate that glucose and fructose share the same membrane transport components.
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    Manganese accumulation in yeast cells (1989)
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    BioMetals 2 (1989), S. 6-10 
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    ISSN: 1572-8773
    Keywords: Manganese ; Electron spin resonance ; Superoxide dismutase ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Manganese accumulation was studied by room-temperature electron spin resonance (ESR) spectroscopy inSaccharomyces cerevisiae grown in the presence of increasing amounts of MnSO4. Mn2+ retention was nearly linear in intact cells for fractions related to both low-molecular-mass and macromolecular complexes (‘free’ and ‘bound’ Mn2+, respectively). A deviation from linearity was observed in cell extracts between the control value and 0.1 mM Mn2+, indicating more efficient accumulation at low Mn2+ concentrations. The difference in slopes between the two straight lines describing Mn2+ retention at concentrations lower and higher than 0.1 mM, respectively, was quite large for the free Mn2+ fraction. Furthermore it was unaffected by subsequent dialyses of the extracts, showing stable retention in the form of low-molecular-mass complexes. In contrast, the slope of the line describing retention of ‘bound’ Mn2+ at concentrations higher than 0.1 mM became less steep after subsequent dialyses of the cell extracts. This result indicates that the macromolecule-bound Mn2+ was essentially associated with particulate structures. In contrast to Cu2+, Mn2+ had no effect on the major enzyme activities involved in oxygen metabolism except for a slight increase of cyanide-resistant Mn-superoxide dismutase activity, due to dialyzable Mn2+ complexes.
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    Cu(I)-thionein release from copper-loaded yeast cells (1989)
    Springer
    BioMetals 2 (1989), S. 50-54 
    Details
    ISSN: 1572-8773
    Keywords: Cu(I)8-thionein ; Yeast ; Extracellular ; Circular dichroism ; Fluorescence ; Electronic absorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The release of intact CU(I)8-thionein from copper-resistant copper-loaded yeast cells, strain X 2180-1Aa, has been shown. This copper(I)-thiolate-rich protein was characterized and compared with the chemical and physicochemical properties of intracellular yeast Cu-thionein. The same molecular mass and stoichiometry of 8 mol copper atoms/mol protein was found. No detectable difference between the Cu-thioneins was seen in luminescence emission, electronic absorption in the ultraviolet region, chiroptical data or amino acid composition. The importance of stable Cu(I)-thiolates in Cu-thionein as a safe vehicle for transporting copper in a non-reactive manner is confirmed.
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    Biotransformation of aromatic aldehydes bySaccharomyces cerevisiae: Investigation of reaction rates (1989)
    Springer
    Journal of industrial microbiology and biotechnology 4 (1989), S. 49-53 
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    ISSN: 1476-5535
    Keywords: l-Phenylacetyl carbinol ; Saccharomyces cerevisiae ; Yeast ; Benzaldehyde ; Biotransformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The rate of production ofl-phenylacetyl carbinol bySaccharomyces cerevisiae in reaction mixtures containing benzaldehyde with sucrose or pyruvate as cosubstrate was investigated in short 1 h incubations. The effect of yeast dose rate, sucrose and benzaldehyde concentration and pH on the rate of reaction was determined. Maximum biotransformation rates were obtained with concentrations of benzaldehyde, sucrose and yeast of 6 g, 40 g and 60 g/l, respectively. Negligible biotransformation rates were observed at a concentration of 8 g/l benzaldehyde. The reaction had a pH optimum of 4.0–4.5. Rates of bioconversion of benzaldehyde and selected substituted aromatic aldehydes using both sucrose and sodium pyruvate as cosubstrate were compared. The rate of aromatic alcohol production was much higher when sucrose was used rather than pyruvate.o-Tolualdehyde and 1-chlorobenzaldehyde were poor substrates for aromatic carbinol formation although the latter produced significant aromatic alcohol in sucrose-containing media. Yields of 2.74 and 3.80 g/l phenylacetyl carbinol were produced from sucrose and pyruvate, respectively, in a 1 h reaction period.
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    Pores from mitochondrial outer membranes of yeast and a porin-deficient yeast mutant: A comparison (1989)
    Springer
    Journal of bioenergetics and biomembranes 21 (1989), S. 439-450 
    Details
    ISSN: 1573-6881
    Keywords: Yeast ; yeast mutant ; mitochondrial porin ; mitochondrial outer membrane ; lipid bilayer ; ion-channel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Reconstitution experiments were performed on lipid bilayer membranes in the presence of purified mitochondrial porin from yeast and of detergent-solubilized mitochondrial outer membranes of a porin-free yeast mutant. The addition of the porin resulted in a strong increase of the membrane conductance, which was caused by the formation of ion-permeable channels in the membranes. Yeast porin has a single-channel conductance of 4.2 nS in 1 M KCl. In the open state it behaves as a general diffusion pore with an effective diameter of 1.7 nm and possesses properties similar to other mitochondrial porins. Surprisingly, the membrane conductance also increased in the presence of detergent extracts of the mitochondrial outer membrane of the mutant. Single-channel recordings of lipid bilayer membranes in the presence of small concentration of the mutant membranes suggested that this membrane also contained a pore. The reconstituted pores had a single-channel conductance of 2.0 nS in 1 M KCl and the characteristics of general diffusion pores with an estimated effective diameter of 1.2 nm. This means that the pores present in the mitochondrial outer membranes of the yeast mutant have a much smaller effective diameter than “normal” mitochondrial porins. Zero-current membrane potential measurements suggested that the second mitochondrial porin is slightly cation-selective, while yeast porin is slightly anion-selective in the open state but highly cation-selective in the closed state. The possible role of these pores in the metabolism of mitochondria is discussed.
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    Isolation of the RAD18 gene of Saccharomyces cerevisiae and construction of rad18 deletion mutants (1989)
    Springer
    Molecular genetics and genomics 215 (1989), S. 425-430 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; DNA repair ; RAD18 ; Chromosomal deletions ; Mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The RAD18 gene of Saccharomyces cerevisiae is involved in mutagenic DNA repair. We describe its isolation from a yeast library introduced into the centromeric YCp50 vector, a low copy number plasmid. The insert was sublconed into YCp50 and into the multicopy YRp7 plasmid. RAD18 is not toxic when present in multiple copies but the UV survival response indicates an heterogeneity in the cell population, a fraction of it being more sensitive. A DNA segment, close to RAD18, is toxic on the multicopy plasmid and may correspond to the tRAN sup61 known to be tightly linked to RAD18. Chromosomal deletions of RAD18 were constructed. The gene is not essential and the deleted strains have the properties of single site mutants. Thus, RAD18 appears to be essentially involved in DNA repair metabolism.
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    Novel class of nuclear genes involved in both mRNA splicing and protein synthesis in Saccharomyces cerevisiae mitochondria (1989)
    Springer
    Molecular genetics and genomics 215 (1989), S. 517-528 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Nuclear genes ; Mitochondrial translation ; Mitochondrial splicing ; Suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned three distinct nuclear genes, NAM1, NAM7, and NAM8, which alleviate mitochondrial intron mutations of the cytochrome b and COXI (subunit I of cytochrome oxidase) genes when present on multicopy plasmids. These nuclear genes show no sequence homology to each other and are localized on different chromosomes: NAM1 on chromosome IV, NAM7 on chromosome XIII and NAM8 on chromosome VIII. Sequence analysis of the NAM1 gene shows that it encodes a protein of 440 amino acids with a typical presequence that would target the protein to the mitochondrial matrix. Inactivation of the NAM1 gene by gene transplacement leads to a dramatic reduction of the overall synthesis of mitochondrial protein, and a complete absence of the COXI protein which is the result of a specific block in COXI pre-mRNA splicing. The possible mechanisms by which the NAM1 gene product may function are discussed.
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    A functional analysis of the repeated methionine initiator tRNA genes (IMT) in yeast (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 276-286 
    Details
    ISSN: 1617-4623
    Keywords: Methionine ; Initiator tRNA ; tRNA(met) ; Yeast ; Multigene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Standard laboratory yeast strains have from four to five genes encoding the methionine initiator tRNA (IMT). Strain S288C has four IMT genes with identical coding sequences that are colinear with the RNA sequence of tRNA I Met . Each of the four IMT genes from strain S288C is located on a different chromosome. A fifth IMT gene with the same coding sequence is present in strain A364A but not in S288C. By making combinations of null alleles in strain S288C, we show that each of the four IMT genes is functional and that tRNA I Met is not limiting in yeast strains with three or more intact genes. Strains containing a single IMT2, 3 or 4 gene grow only after amplification of the remaining IMT gene. Strains with only the IMT1 gene intact are viable but grow extremely slowly; normal growth is restored by the addition of another IMT gene by transformation, providing a direct test for IMT function.
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    α-Factor leader sequence-directed transport of Escherichia coli β-galactosidase in the secretory pathway of Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 218 (1989), S. 240-248 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; MFα1 leader ; Gene fusion ; Secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The constuction of two fused genes is described. One involves the in-frame fusion of the yeast prepro-α-factor coding sequence, and the Escherichia coli lac Z gene. The second gene fusion utilizes a 103 bp yeast invertase NH2-terminal coding sequence at the fusion junction of the hybrid gene described above. The gene fusions, under the control of the α-factor promoter, expressed active β-galactosidase in α haploid yeast cells. The activity could be regulated in a temperature-sensitive sir3 mutant. The incorporation of the invertase coding sequence at the MFα1-lacZ fusion junction provided significantly higher levels of β-galactosidase activity. A substantial quantity of the hybrid proteins generated from the gene fusions was primarily localized in the intracellular membranes of yeast cells, while a processed form could be secreted into the periplasm.
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    Mode of expression of the positive regulatory genes PHO2 and PHO4 of the phosphatase regulon in Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 31-39 
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    ISSN: 1617-4623
    Keywords: Autoregulation ; LacZ fusion protein ; Northern hybridization ; Regulatory circuit ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mode of expression was investigated for two positive regulatory genes PHO2 and PHO4, whose products are indispensable for the transcriptional control of the structural genes of repressible acid phosphatase and the inorganic phosphate (Pi) transport system in Saccharomyces cerevisiae. Northern analysis of poly(A)+ RNA of the wild-type and the pho regulatory mutants with PHO4 DNA as hybridization probe and expressional analysis of a pho4′-'lacZ fused gene on a YEp plasmid revealed that PHO4 is expressed at a low level, constitutively, and independently of the PHO regulatory system and Pi in the medium. Similar analyses with PHO2 DNA indicated that PHO2 is expressed at an even lower level than PHO4, and is repressed by Pi and by the active PHO2 product, possibly at the translational level, while retaining a substantial level of basal activity.
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    Function of the PHO regulatory genes for repressible acid phosphatase synthesis in Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 40-46 
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    ISSN: 1617-4623
    Keywords: Gene dosage ; Gene expression ; Regulatory circuit ; Signal transmission ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Expression of the repressible acid phosphatase (rAPase) gene, PHO5, of Saccharomyces cerevisiae is repressed by a certain level of inorganic phosphate (Pi) in the medium and is derepressed when the Pi concentration is lowered. The Pi signals are conveyed to PHO5 by a regulatory system consisting of proteins coded for by the PHO2, PHO4, PHO80 and PHO81 genes. We have found that the transcription of PHO81 is regulated by Pi through the PHO regulatory system. Increasing the dosage of PHO4 and PHO81 by ligating each gene to YEP13 gives rise to, respectively, considerable and weak synthesis of rAPase by cultivation of the transformants in high-Pi medium; but in low-Pi medium, increased dosage of PHO4 stimulates the rAPase synthesis significantly, whereas PHO81 has no effect. Increased dosage of PHO2 stimulates rAPase synthesis considerably in low-Pi but not in high-Pi. A coordinate increase of PHO80 cancels the dosage effect of PHO4, but not that of PHO81. Coordinate increases of PHO80 and PHO2 give rise to the same phenotype as an increased dosage of PHO80 alone. The level of the PHO4 protein was found to be the limiting factor of the rAPase synthesis and the copy number of the PHO5 gene not to be. These facts accord with the idea that the PHO80 protein transmits the Pi signals to the PHO5 gene via the PHO4 protein, whereas the PHO2 protein does not have a direct function in the signal transmission.
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    The paromomycin resistance mutation (par r-454) in the 15 S rRNA gene of the yeastSaccharomyces cerevisiae is involved in ribosomal frameshifting (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 362-369 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondrial 15 S rRNA ; Ribosomal frameshifting ; Paromomycin resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The leaky expression of the yeast mitochondrial geneoxi1, containing a frameshift mutation (+1), is caused by natural frameshift suppression, as shown previously (Fox and Weiss-Brummer 1980). A drastic decrease in the natural level of frameshifting is found in the presence of thepar r-454 mutation, localized at the 3′ end of the 15 S rRNA gene. This mutation causes resistance to the antibiotic paronomycin in the yeast strains D273-10B and KL14-4A (Li et al. 1982; Tabak et al. 1982). The results of this study imply that in the yeast strain 777-3A this mutation alone is sufficient for restriction of the level of natural frameshifting but is insufficient to confer resistance to paromomycin. A second mutation, arising spontaneously with a frequency of 10−4 leads, in combination with thepar r-454 mutation, to full paromomycin resistance in strain 777-3A.
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    Chromatin digestion with restriction endonucleases reveals 150–160 bp of protected DNA in the centromere of chromosome XIV in Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 153-160 
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    ISSN: 1617-4623
    Keywords: Centromere ; Chromatin ; Hypersensitive sites ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Isolated nuclei of Saccharomyces cerevisiae were incubated with five restriction nucleases. Out of the twenty-one recognition sequences for these nucleases in the centromere region of chromosome XIV, only five are accessible to cleavage. These sites map 11 by and 74 by to the left and 27 bp, 41 by and 290 by to the right, respectively, of the boundaries of the 118 by functional CEN14 DNA sequence. The distance between the sites accessible to cleavage and closest to CEN14 is 156 bp, suggesting this is the maximal size of DNA protected in CEN14 chromatin. The DNA in CEN14 chromatin protected against cleavage with DNase I and micrococcal nuclease overlaps almost completely with this region. Hypersensitive regions flanking both sides are approximately 60 by long. Analyses of other S. cerevisiae centromeres with footprinting techniques in intact cells or nucleolytic cleavages in isolated nuclei are discussed in relation to our results. We conclude that structural data of chromatin obtained with restriction nucleases are reliable and that the structure of CEN14 chromatin is representative for S. cerevisiae centromeres.
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    Molecular characterization of a specific p-nitrophenylphosphatase gene, PHO13, and its mapping by chromosome fragmentation in Saccbaromyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 220 (1989), S. 133-139 
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    ISSN: 1617-4623
    Keywords: Chromosome fragmentation ; Mapping ; PHO13 sequence ; Phosphatase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The structural gene, PHO13, for the specific p-nitrophenyl phosphatase of Saccharomyces cerevisiae was cloned and its nucleotide sequence determined. The deduced PHO13 protein consists of 312 amino acids and its molecular weight is 34635. The disruption of the PHO13 gene produced no effect on cell growth, sporulation, or viability of ascospores. The PHO13 locus was mapped at 1.9 centimorgans from the HO locus on the left arm of chromosome IV. By chromosome fragmentation, the PHO13 locus was found to be located about 72 kb from the left-hand telomere of chromosome IV and distal to the HO locus.
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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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    How hexoses and inhibitors influence the malate transport system in Zygosaccharomyces bailii (1988)
    Springer
    Archives of microbiology 150 (1988), S. 37-41 
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    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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    Chloroquine, a lysosomotropic agent, inhibits zygote formation in yeast (1988)
    Springer
    Archives of microbiology 151 (1988), S. 20-25 
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    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.
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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 
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    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.
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    Interspecific protoplast fusion between the yeasts Saccharomyces cerevisiae and Saccharomyces fermentati (1988)
    Springer
    Current genetics 13 (1988), S. 101-104 
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    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.
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    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 
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    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.
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    Induced recombination and reversion of theCDC8 gene in relation to the cell cycle of yeast (1988)
    Springer
    Current genetics 13 (1988), S. 455-460 
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    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.
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    Modifiers of ochre suppressors in Saccharomyces cerevisiae that exhibit ochre suppressor-dependent amber suppression (1988)
    Springer
    Current genetics 14 (1988), S. 345-354 
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    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.
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    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 
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    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.
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    Multiple elements regulate expression of the cell cycle-regulated thymidylate synthase gene of Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 363-373 
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    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.
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    Immunological homologies between yeast ribosomal protein L2 and rat liver ribosomal proteins L4 and L24 (1988)
    Springer
    Current genetics 13 (1988), S. 235-239 
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    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.
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    Plasmid associations with residual nuclear structures in Saccharomyces cerevisiae (1988)
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    Current genetics 13 (1988), S. 291-297 
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    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.
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    FLP recombinase induction of the breakage-fusion-bridge cycle and gene conversion in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 273-281 
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    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.
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    A reexamination of the role of the RAD52 gene in spontaneous mitotic recombination (1988)
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    Current genetics 14 (1988), S. 211-223 
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    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.
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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 
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    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.
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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 
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    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.
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    Sensitivity of yeast glycolytic enzymes to chloroquine (1988)
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    Archives of microbiology 150 (1988), S. 309-312 
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    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).
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    Thermotolerant single cell protein production byKluyveromyces marxianus var.marxianus (1988)
    Springer
    Journal of industrial microbiology and biotechnology 3 (1988), S. 9-14 
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    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.
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    Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation (1988)
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    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
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    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.
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    Carbon assimilation tests: Substrates assimilation profiles (1988)
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    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.
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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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    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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    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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    Rad3 protein of Saccharomyces cerevisiae: Overexpression and preliminary characterization using specific antibodies (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 400-408 
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    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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    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.
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    URL: http://dx.doi.org/10.1007/BF00340176
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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 
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    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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    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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    The structure and regulation of phosphoglucose isomerase in Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 100-106 
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    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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    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 
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    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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    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 
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    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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    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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    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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    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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    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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    Molecular evolution of theSaccharomyces cerevisiae histone gene loci (1987)
    Springer
    Journal of molecular evolution 24 (1987), S. 252-259 
    Details
    ISSN: 1432-1432
    Keywords: Histone genes ; Gene conversion ; Diploidization ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The core histone genes ofSaccharomyces cerevisiae are arranged as duplicate nonallelic sets of specifically paired genes. The identity of structural organization between the duplicated gene pairs would have its simplest evolutionary origin in the duplication of a complete locus in a single event. In such a case, the time since the duplication of one of the genes should be identical to that since duplication of the gene adjacent to it on the chromosome. A calculation of the evolutionary distances between the coding DNA sequences of the histone genes leads to a duplication paradox: The extents of sequence divergence in the silent component of third-base positions for adjacent pairs of genes are not identical. Estimates of the evolutionary distance between the two H3-H4 noncoding intergene DNA sequences are large; the divergence between the two separate sequences is indistinguishable from the divergence between either of the regions and a randomly generated permutation of itself. These results suggest that the duplication event may have occurred much earlier than previously estimated. The potential age of the duplication, and the attractive simplicity of the duplication of both the H3-H4 and the H2A-H2B gene pairs having taken place in a single event, leads to the hypothesis that modern haploidS. cerevisiae may have evolved by diploidization or fusion of two ancient fungi.
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    Yeast centromere sequences do not confer mitotic stability on circular plasmids containing ARS elements of Tetrahymena thermophila rDNA (1987)
    Springer
    Current genetics 11 (1987), S. 353-357 
    Details
    ISSN: 1432-0983
    Keywords: Tetrahymena ; Replication ; Segregation ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have previously demonstrated that a 657 bp TaqI-XbaI and a 427 by XbaI-XbaI fragment from the 5′ non-transcribed spacer of the extrachromosomal ribosomal DNA of Tetrahymena thermophila function as autonomously replicating sequences (ARS) in Saccharomyces cerevisiae. These fragments are adjacent to each other in a region that encompasses the in vivo origin of bidirectional replication of rDNA. The presence of a yeast centromere (CEN) fragment does not confer mitotic stability on these plasmids. A sensitive yeast colony colour assay (Hieter et al. 1985a) has been used to evaluate the cis-acting effect of each ARS segment on the pattern of inheritance of a plasmid containing CEN5:URA3:SUP4. Colonies of transformed cells obtained both in the presence and absence of selection were red with no detectable white or pink sectors. The lack of sectoring indicates that both plasmids are lost at an extremely high rate, likely due to 1:0 segregation events. We conclude that while these ARS elements confer a high frequency transformation phenotype, they lack a function which is required in cis for the maintenance of mitotic stability in the presence of a centromere. This missing cis-acting function may result in the inability of the plasmids to be brought under the control of cell-cycle regulated replication.
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    Temperature sensitive pet mutants in yeast Saccharomyces cerevisiae that lose mitochondrial RNA (1987)
    Springer
    Current genetics 11 (1987), S. 359-367 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial ; Mutants ; RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This is a description of a new class of temperature sensitive pet mutants in Saccharomyces cereviase that lose all or part of their mitochondrial RNA at the restrictive temperature. These mutants fall into 8 different complementation groups, mna1 to mna8, and 2 different classes based on their phenotype. Class I mutations, mna1-1 through mna5-1, cause complete or partial loss of mitochondrial RNA at the restrictive temperature. The mutation, mna1-1, is especially interesting since it causes a loss of both mitochondrial DNA and RNA when the mutant is grown on a fermentable carbon source at the restrictive temperature. However, when this mutant is grown at the permissive temperature on a non-fermentable carbon source then shifted to the restrictive temperature, only the mitochondrial RNA is lost. This indicates that the primary cause for the pet phenotype is due to the loss of mitochondrial RNA and not DNA. Class II mutations, mna6-1 through man8-1, cause complete loss of the 14S rRNA after growth at the restrictive temperature in a fermentable carbon source. This loss appears to be specific for the 14S rRNA, since all other transcripts probed by Northern analysis are normal.
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    A mitochondrial frameshift-suppressor (+) of the yeast S. cerevisiae maps in the mitochondrial 15S rRNA locus (1987)
    Springer
    Current genetics 11 (1987), S. 295-301 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial ; Frameshift-Suppression ; 15S rRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The first case of a +1 “extrageneic” frameshift suppressor (MF1), mapping in the yeast mitochondrial 15S rRNA gene is reported. The suppressor was identified by genetic analyses in a leaky mitochondrial oxi1 frameshift mutant and the respective wild-type strain 777-3A of the yeast S. cerevisiae. This is in accordance with the finding that all mitochondrial frameshift mutants isolated from this strain tend to be leaky to a variable degree. MF1 does not suppress known nonsense mutations created by a direct basepair exchange in strain 777-3A. These mutants exhibit a non-leaky phenotype (Weiss-Brummer et al. 1984).
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    The Yarrowia lipolytica LEU2 gene (1987)
    Springer
    Current genetics 11 (1987), S. 377-383 
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    ISSN: 1432-0983
    Keywords: Y. lipolytica ; LEU2 ; Yeast ; Leucine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2810 by DNA fragment containing the beta-isopropylmalate dehydrogenase gene of the dimorphic yeast Yarrowia lipolytica has been sequenced. The sequence contains an open reading frame of 405 codons, predicting a protein of 43,366 molecular weight. Protein sequence homology with the polypeptide encoded by the LEU2 gene of Saccharomyces cerevisiae is 64%, whereas DNA sequence homology is 61%. The 5′- and 3′-flanking regions of the Y. lipolytica LEU2 gene share only some general structural features common to genes of S. cereviside such as the presence and location of TATA boxes, CAAT boxes, CACACA repeats, the lack of G residues in the 5′-untranslated region and 3′-transcription terminators. Transcription of a 1.4 kb mRNA begins at a small cluster of sites approximately 40 base pairs before the initial ATG.
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    Genetic modification of the spontaneous rho − mutability in Saccharomyces cerevisiaee (1987)
    Springer
    Current genetics 11 (1987), S. 411-413 
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    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial rho − mutability ; Genetic analysis ; Modifying genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The phenotypic trait “starry colony” in Saccharomyces is associated with a high spontaneous rho − petite mutability. Genetic analysis of this trait has shown the high rho − mutability to be caused by several modifying genes present together in the strains studied. Every single modifying gene produces only a relatively small enhancement of the rho − mutability.
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    Base analogue mutagenesis in yeast and its modulation by pyrimidine deoxynucleotide pool imbalances: incorporation of bromodeoxyuridylate and iododeoxyuridylate (1987)
    Springer
    Current genetics 11 (1987), S. 421-427 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mutagenesis ; Base analogues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cells of the yeast, Saccharomyces cerevisiae, which are auxotrophic for thymidylate (tmp1) can also incorporate analogues of thymidylate. When the base analogue, 5-bromodeoxyuridylate, is incorporated into tmp1 yeast cells it is lethal and mutagenic. Both lethality and mutation induction can be drastically altered by perturbation of the pyrimidine nucleotide pools. Analysis of mutation induction, bromodeoxyuridylate incorporation into DNA, and cell viability under various conditions revealed: (1) lethality and mutagenesis can be uncoupled, (2) thymidylate enhances mutagenesis and deoxycytidylate suppresses it, (3) mutation induction is not correlated with the magnitude of bromodeoxyuridylate incorporation into DNA. Therefore, in yeast, the pyrimidine nucleotide pools have a powerful effect on bromodeoxyuridylate mutagenesis. Both bromodeoxyuridylate and iododeoxyuridylate are extensively incorporated into the DNA of tmp1 yeast cells; however, iododeoxyuridylate is non-mutagenic. Replication proceeds at the same rate in the presence of the natural substrate or either analogue. When cells are supplied with thymidylate and bromodeoxyuridylate together, there is no discrimination against bromodeoxyuridylate as a DNA precursor. However, in the presence of thymidylate and iododeoxyuridylate, there is a 3 to 1 discrimination against iododeoxyuridylate as compared to thymidylate.
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    URL: http://dx.doi.org/10.1007/BF00384602
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    Extrachromosomal genetics in the yeast Kluyveromyces lactis. Isolation and characterization of antimycin-resistant mutants (1987)
    Springer
    Current genetics 11 (1987), S. 475-482 
    Details
    ISSN: 1432-0983
    Keywords: Kluyveromyces lactis ; Yeast ; Extrachromosomal inheritance ; Antimycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Antimycin-resistant (AR) mutants of the yeast Kluyveromyces lactis, obtained either spontaneously or after manganese treatment, were isolated and genetically characterized. Most of the mutants obtained after manganese mutagenesis and two spontaneous mutants, tolerated high antimycin concentrations (more than 10 /gmg/ml) and were extrachromosomal. One mutant which grew only in low antimycin (1 /gmg/ml) showed a Mendelian type of inheritance. The extrachromosomal mutants could be assigned to at least two genetic loci (A I R and A II R ). Mutants representative of these two groups showed increased resistance to the antibiotic when the respiration of whole cells or mitochondria was studied. Extrachromosomal mutants of Saccharomyces cerevisiae resistant to antimycin were also induced with manganese, isolated and characterized. Comparative studies of the antimycin-resistant mutants of K. lactis and S. cerevisiae permitted the following observations: a) K. lactis is more resistant to antimycin, funiculosin, mucidin and diuron than S. cerevisiae, as are the AR mutants; b) K. lactis shows correlated sensitivity to funiculosin differing in this aspect from S. cerevisiae; c) the antimycin-resistant mutants of K. lactis belonging to group 11 (A II R ) were also resistant to diuron, tolerating concentrations of more than 200 /gmg/ml; d) all extrachromosomal antimycin-resistant-mutants of S. cerevisiae and some of the AR mutants of K. lactis were more sensitive to mucidin than the wild type.
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    URL: http://dx.doi.org/10.1007/BF00384609
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    Cryptic DNA plasmids of the heterothallic yeast Saccharomycopsis crataegensis (1987)
    Springer
    Current genetics 12 (1987), S. 297-304 
    Details
    ISSN: 1432-0983
    Keywords: Fungi ; S. crataegensis ; Yeast ; Plasmid ; Linear DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three DNA plasmids, designated pScrl-1, pScrl-2, and pScrl-3 have been found in a strain of the heterothallic yeast Saccharomycopsis crataegensis (NRRL Y-5902). pScrl-l, -2 and -3 are, respectively, 15, 7, and 5 kilobase pairs (kbp) in size. Based on the results of exonuclease digestions, all three plasmids appear to be linear molecules with blocked 5′ ends. All three plasmids also have a lower buoyant density than does nuclear DNA of S. crataegensis. The two lower molecular weight plasmids hybridize strongly with one another, but only weakly to the higher molecular weight plasmid. Two of four related S. crataegensis strains surveyed were found to contain two plasmids that are of the same size as the two larger plasmids of Y-5902. Evidence is presented indicating that the plasmids in strain Y-5902 reside in the cytosol since they were found not to be located within the major organelles (mitochondria and nuclei).
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    URL: http://dx.doi.org/10.1007/BF00435293
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  • 86
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    At least two nuclear-encoded factors are involved together with a mitochondrial factor (MR1) in spontaneous mitochondrial frameshift-suppression of the yeast S. cerevisiae (1987)
    Springer
    Current genetics 12 (1987), S. 387-390 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Frameshift-Suppression ; Mitochondrial/Nuclear ; Interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Earlier genetic analyses have identified a mitochondrial +1 frameshift suppressor (MF1) in the 15S rRNA region of a leaky mitochondrial frameshift mutant and the respective wild-type strain 777-3A (Weiss-Brummer et al. 1987). Further genetic analyses revealed that for the observed spontaneous frameshift suppression in M5631 the mitochondrial factor (MF1) must act together with at least two dominant nuclear-encoded factors.
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    URL: http://dx.doi.org/10.1007/BF00405761
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  • 87
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    Phenotypic suppression and nuclear accommodation of the mit − oxi1-V25 mutation in isolated yeast mitochondria (1987)
    Springer
    Current genetics 12 (1987), S. 305-310 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Translation ; Informational Suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Phenotypic suppression by the antibiotic, paromomycin, of the mitochondrial oxi1 −-V25 mutation, a mutation which arrests by premature ochre codon the synthesis of the cox 11 subunit, was studied in isolated yeast mitochondria competent in translation. This antibiotic is known to suppress the mutation in vivo (Dujardin et al. 1984) and allowed in vitro, at concentrations of 20–1100 Mg per ml. the synthesis of the cox II subunit. This strongly suggests that phenotypic suppression of mit − mutations is due to the direct action of paromomycin on mitochondrial ribosomes. The effect of paromomycin bears a resemblance to the function of the omnipotent nuclear suppressor mutation R705. The nuclear suppression was expressed in isolated mitochondria; suppressor mutation influenced the structure of the mitoribosome. Therefore, it appears that mitoribosomes are indeed the common target in the phenotypical and genetic nuclear suppression of the oxi1-V25 mutation.
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    URL: http://dx.doi.org/10.1007/BF00405752
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  • 88
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    The disomy for chromosome IV and the spontaneous rho- mutability in Saccharomyces cerevisiae (1987)
    Springer
    Current genetics 11 (1987), S. 407-410 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Disomy for chromosome IV ; Mitochondrial rho − mutability ; Mitotic chromosome loss
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The disomy for chromosome IV in the strains studied led to: i) a reduction in the red pigmentation of ade1 mutant colonies; ii) a decrease of the spontaneous rho − mutant frequency, and iii) an impairment of sporulation in hybrids descended from disomic parents. The nuclear srm1 mutation decreasing the spontaneous rho − mutability promoted the spontaneous extra chromosome loss in the disomes for chromosome IV. This result suggests a close connexion between the spontaneous rho − mutability and mitotic chromosome stability.
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    URL: http://dx.doi.org/10.1007/BF00378184
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  • 89
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    Effect of ploidy on the critical size for cell proliferation of the yeast Saccharomyces cerevisiae (1987)
    Springer
    Current genetics 11 (1987), S. 591-594 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Nuclear ploidy ; Critical size ; Cell proliferation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary For a polyploid series of Saccharomyces cerevisiae strains ranging from haploid to tetraploid we found that the critical cell size required to initiate a new cell division process was directly and linearly proportional to ploidy, but was not influenced by the information at the MAT locus which determines cell type. Therefore, over at least a four-fold range in ploidy the cell cycle machinery which is responsive to growth is modulated by nuclear DNA content.
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    URL: http://dx.doi.org/10.1007/BF00393920
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    Comparative enzyme and ethanol production in an isogenic yeast ploidy series (1987)
    Springer
    Current genetics 12 (1987), S. 9-14 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Ploidy ; Isogenic ; Ethanol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Effects on ethanol production by increases in gene dosage independent of heterosis in yeast are compared for an isogenic ploidy series ranging from haploid to tetraploid. The per-cell rate of ethanol accumulation in parallel batch cultures increases with cell ploidy, and is attributable to intrinsic, ploidy-associated increases in cell mass-adjusted ethanol production rates. This increase in per-cell ethanol accumulation in the tetraploid strain is as high as 6.9 times the level of accumulation in the haploid. That is, the efficiency of ethanol production per unit cell mass is greater in cells of higher ploidy.
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    URL: http://dx.doi.org/10.1007/BF00420721
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    The influence of cell ploidy on the thermotolerance of Saccharomyces cerevisiae (1987)
    Springer
    Current genetics 11 (1987), S. 595-598 
    Details
    ISSN: 1432-0983
    Keywords: Heat shock ; Thermotolerance ; Ploidy ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The resistance of Saccharomyces cerevisiae to inactivation by DNA damaging agents has long been known to be affected by cell ploidy. Resistance is greater for diploid than for haploid cells, but exhibits decreases for further increases in ploidy beyond diploid. In this study S. cerevisiae cells whose genomes differ only in their ploidy were employed to investigate how ploidy directly influences resistance to thermal killing. In virtually all species resistance to thermal killing is a cellular property that is elevated by heat shock and other agents that induce the heat shock response. We therefore investigated how ploidy affected the thermal killing of S. cerevisiae cells both before and after elevation of thermotolerance by means of a 40 min 25 °C to 38 °C heat shock. Without such induction of thermotolerance there was negligible effect of ploidy on thermal killing. In contrast in the heat shocked cultures there was an appreciable decrease in thermotolerance as ploidy increased. This difference indicates that the lethal thermal damage in the thermotolerance induced cultures is not totally equivalent to that in cells not given a prior heat shock, and that gene expression changes after heat shock result in a ploidy effect on heat tolerance which is absent from cells in which the heat shock response has not been induced.
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    URL: http://dx.doi.org/10.1007/BF00393921
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    Coincident chromosomal disomy in meiotic dyads from triploid yeast (1987)
    Springer
    Current genetics 12 (1987), S. 569-576 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Disomy ; Meiotic dyads
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Among meiotic asci produced by triploid (3N) Saccharomyces cerevisiae are cases in which exactly two of the four ascospores proliferate into colonies. Given the unique asymmetry problems inherent in distributing three chromosome homologues in meiosis, these ascospore dyads are of special interest. We have tested 40 of these dyads (80 ascospores) for their chromosome content by ascertaining whether they have inherited one or two copies of each of the sixteen yeast chromosomes from the parental triploid. Overall, then, ascospores in these dyads can be either haploid (N) or disomic (N + 1) for each chromosome. The principal results of this analysis include: (1) Coincident disomy (inheritence of two copies of a given chromosome in both members of an ascospore dyad) was detected for 15 of the 16 yeast chromosomes, and at least once in every dyad. (2) Coincident disomy increased as a function of the mean number of disomic chromosomes per spore in each dyad, but this increase differed functionally from that expected if coincident disomy in the two ascospores were a simple, meiotically independent, concomitant of multiple disomy. We conclude from these results that: (1) The ascospore dyads, as the two proliferating spores of single meioses from the triploid, represent meiotic sisters. That is, they stem from the same half of the first meiotic division. (2) Multiply-disomic meiotic segregants of yeast triploids proliferate at the expense of their multiple disomy, as cells in spore colonies experience repeated and independent disomic chromosome losses (N + 1 → N). (3) Aneuploid generation in triploid meiosis is chromosomally unbiased and is the consequence of the independent two-by-one segregation at MI of every homologous triad.
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    URL: http://dx.doi.org/10.1007/BF00368058
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    ARS activity along the yeast mitochondrial apocytochrome b region: correlation with the location of petite genomes and consensus sequences (1987)
    Springer
    Current genetics 12 (1987), S. 583-589 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; ARS-like activity ; Petite genome replication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Seven MboI fragments spanning the mitochondrial apocytochrome b gene in Saccharomyces cerevisiae strain D273-10B were cloned in the BamHI site of the integrative yeast vector YIp5 and the capacity for autonomous replication was subsequently assayed in yeast. The positive correlation found between the ars-like activity in four fragments and the presence of regions common to multiple ethidium bromide-induced petite (rho−) genomes suggests that the mitochondrial sequences possibly active as origins of replication in low-complexity neutral or weakly suppressive rho− mutants could be functionally related to the yeast nuclear replicator 11 nucleotide motif defined by Broach et al. (1983).
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    URL: http://dx.doi.org/10.1007/BF00368060
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    A proton-translocating adenosine triphosphatase is associated with the peroxisomal membrane of yeasts (1987)
    Springer
    Archives of microbiology 147 (1987), S. 42-47 
    Details
    ISSN: 1432-072X
    Keywords: Hansenula polymorpha ; Yeast ; Peroxisomes ; Proton-translocating ATPase ; Cell fractionation ; Fluorescence quenching studies ; Cytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The association of an ATPase with the yeast peroxisomal membrane was established by both biochemical and cytochemical procedures. Peroxisomes were purified from protoplast homogenates of the methanol-grown yeast Hansenula polymorpha by differential and sucrose gradient centrifugation. Biochemical analysis revealed that ATPase activity was associated with the peroxisomal peak fractions which were identified on the basis of alcohol oxidase and catalase activity. The properties of this ATPase closely resembled those of the mitochondrial ATPase of this yeast. The enzyme was Mg2+-dependent, had a pH optimum of approximately 8.5 and was sensitive to N,N′-dicyclohexylcarbodiimide (DCCD), oligomycin and azide, but not to vanadate. A major difference was the apparent K m for ATP which was 4–6 mM for the peroxisomal ATPase compared to 0.6–0.9 mM for the mitochondrial enzyme. Cytochemical experiments indicated that the peroxisomal ATPase was associated with the membranes surrounding these organelles. After incubations with CeCl3 and ATP specific reaction products were localized on the peroxisomal membrane, both when unfixed isolated peroxisomes or formaldehyde-fixed protoplasts were used. This staining was strictly ATP-dependent; in controls performed i) in the absence of substrate, ii) in the presence of glycerol 2-phosphate instead of ATP, or iii) in the presence of DCCD, staining was invariably absent. Similar staining patterns were observed in subcellular fractions and protoplasts of Candida utilis and Trichosporon cutaneum X4, grown in the presence of ethanol/ethylamine or ethylamine, respectively.
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    Effect of ozone on ATP, cytosolic enzymes and permeability of Saccharomyces cerevisiae (1987)
    Springer
    Archives of microbiology 147 (1987), S. 105-108 
    Details
    ISSN: 1432-072X
    Keywords: Ozone ; Yeast ; Saccharomyces cerevisiae ; ATP ; Nucleotides ; Permeability ; Cytosolic enzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Treatment of a yeast suspension with ozone inactivates a number of cytosolic enzymes. Among 15 studied, the most drastic inactivation was found for glyceraldehyde-3-phosphate dehydrogenase and to lesser extents: NAD-glutamate dehydrogenase, pyruvate decarboxylase, phosphofructokinase-1 and NAD-alcohol dehydrogenase. Ozone treatment also effects the quantity of ATP and of other nucleoside triphosphates, reducing to about 50% of the initial value. The ATP missing in the cells appears in the medium. NAD and protein also accumulate in the medium suggesting that the yeast cells have been permeabilized. Permeabilization of the yeast cells by treatment with ozone preceeds the inactivation of glyceraldehyde-3-phosphate dehydrogenase and other cytosolic enzymes.
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    URL: http://dx.doi.org/10.1007/BF00415269
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    Inhibition of protein phosphorylation by chloroquine (1987)
    Springer
    Archives of microbiology 147 (1987), S. 235-239 
    Details
    ISSN: 1432-072X
    Keywords: Chloroquine ; Yeast ; Fructose-1,6-bisphosphatase ; Phosphorylation ; Protein kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The rapid phase of fructose-1,6-bisphosphatase (FBPase) inactivation following glucose addition to starved yeast cells [reported previously] is inhibited on addition of 10 mM chloroquine (CQ) at about pH 8. This inhibition of inactivation was shown to be due to the prevention of phosphorylation of the enzyme. CQ was also found to inhibit general protein phosphorylation in the yeast cells. Glycolysis, as observed by changes in intracellular glucose-6-phosphate and extracellular glucose and ethanol concentrations, was shown to be significantly inhibited in cells treated with CQ. Similarly, a decrease in ATP concentrations was observed. However, during the early stages of phosphorylation of FBPase, levels of ATP were similar in cells containing CQ as in those without CQ. Thus, decrease in ATP levels is not thought to be significantly responsible for the inhibition of protein phosphorylation. However, the phosphorylating activity of cyclic AMP-dependent protein kinases is inhibited in vitro by relatively low concentrations of CQ. Thus, prevention of protein phosphorylation by CQ is believed to be due to inhibition of protein kinases in yeast cells.
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    URL: http://dx.doi.org/10.1007/BF00463481
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    A quantitative method for predicting shelf life of soft drinks using a model system (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 59-62 
    Details
    ISSN: 1476-5535
    Keywords: Yeast ; Zygosaccharomyces ; Spoilage ; Synergism ; pH ; °Brix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A quantitative method for the prediction of growth of the food spoilage yeastZygosaccharomyces bailii in a model fruit-drink system is described. A factorially designed experiment was employed to produce polynomial equations relating pH and sugar concentration (°brix) to the lag period and doubling time of this yeast. Low pH values (〈3.0) and high °brix values (〉40) show a strong synergistic action on the extension of lag period, which could be used, along with the model presented, in the formulation of product preservation systems.
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    URL: http://dx.doi.org/10.1007/BF01569407
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    Selection in yeast I: Assessing genetic stability and relative fitness of commercial yeasts (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 159-165 
    Details
    ISSN: 1476-5535
    Keywords: Yeast ; Genetic stability ; Saccharomyces cerevisiae ; Selection ; Reproductive fitness
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The potential for changes in allele frequencies in yeast populations by selection was examined. Cells from the wine yeastSaccharomyces cerevisiae (strain Montrachet) were grown over a large number of generations using two different culturing techniques, each with two variations: serial transfers on WLN agar plates with and without UV irradiation, and continuous culture in autoclaved and in filter-sterilized grape must. A low frequency of variant isozyme patterns was found in samples taken at the end of the experiment. Growth rates in must and on agar plates were also examined, and it was found that all samples were faster-growing than the original strain, to varying degrees. Applications for the selection system developed are discussed.
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    URL: http://dx.doi.org/10.1007/BF01569423
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    Selection in yeast II: The fitness distribution of new mutations inSaccharomyces cerevisiae and its use in a computer simulation (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 167-174 
    Details
    ISSN: 1476-5535
    Keywords: Selection ; Yeast ; Fitness distribution ; Mutation ; Saccharomyces cerevisiae ; Computer simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The fitness distribution of new mutations inSaccharomyces cerevisiae strain Montrachet was determined for cells on agar irradiated for four periods of time with ultraviolet light. The fitness distributions were obtained by converting a large number of colony diameters into relative fitnesses. The distributions were then used to perform a computer simulation with the purpose of predicting the potential of a stock culture to increase in general fitness through selection, given a frequency and magnitude of mutations.
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    URL: http://dx.doi.org/10.1007/BF01569424
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    High-productivity fermentation process for cultivating industrial microorganisms (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 79-85 
    Details
    ISSN: 1476-5535
    Keywords: Yeast ; Bacteria ; High cell density ; Oxygen transfer ; Heat transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary High-productivity continuous fermentation processes have been developed for the production of important industrial microorganisms in specially designed fermentors.Saccharomyces cerevisiae, Pichia pastoris, Kluyveromyces fragilis, andCandida utilis yeasts have been grown in bench-scale fermentors at cell densities of over 120 g/l, whileEscherichia coli, Bacillus megaterium, Methylomonas sp. andPseudomonas putida bacteria have been cultivated to cell densities of more than 110 g/l. Productivities (g cells per 1 per h) greater than 25 have been achieved in both bench-scale and 1500-liter fermentors with yeasts, and values as high as 55 have been achieved with bacteria in the bench-scale fermentor. The microorganisms were grown on defined media using ammonia for pH control and as nitrogen source. The fermentor, capable of high oxygen and heat transfer rates, was operated at constant volume with continuous feed and product discharge. The high-productivity process reduces fermentor size, media sterilization requirements, and may under some circumstances eliminate waste and recycle streams. It can also be applied to a variety of biological products.
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    URL: http://dx.doi.org/10.1007/BF01569506
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