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  • Yeast  (162)
  • Springer  (162)
  • 2015-2019
  • 1985-1989  (162)
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Year
  • 1
    Electronic Resource
    Electronic Resource
    The cloning and characterization of a RAS gene fromSchizosaccharomyces pombe (1986)
    Springer
    Journal of molecular evolution 23 (1986), S. 41-51 
    Details
    ISSN: 1432-1432
    Keywords: RAS oncogene ; Cloning ; DNA sequence ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned and determined the complete nucleotide sequence of a RAS gene from the yeastSchizosaccharomyces pombe (SP-RAS). The putative RAS protein of 214 amino acids is encoded by two noncontiguous reading frames separated by an intron of 86 bp. The SP-RAS gene product shares extensive homology with the proteins of theSaccharomyces cerevisiae (SC),Dictyostelium, Drosophila, and human RAS genes in its N-terminal region but not in its C-terminal region. The extended C-terminal regions found in the SC-RAS genes have no counterpart in the SP-RAS gene. Thus the RAS genes of these two yeasts are structurally quite distinct. The SP-RAS sequence was expressed in vivo.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02100997
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  • 2
    Electronic Resource
    Electronic Resource
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02111238
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  • 3
    Electronic Resource
    Electronic Resource
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00492903
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  • 4
    Electronic Resource
    Electronic Resource
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00463481
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  • 5
    Electronic Resource
    Electronic Resource
    Biosynthesis regulation of the β-glucosidase produced by a yeast strain transformed by genetic engineering (1986)
    Springer
    Archives of microbiology 146 (1986), S. 115-117 
    Details
    ISSN: 1432-072X
    Keywords: β-Glucosidase ; Yeast ; Genetic engineering ; Biosynthesis regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The biosynthesis of the β-glucosidase enzyme was studied in a transformed yeast obtained by cloning in Saccharomyces cerevisiae the structural gene coding for β-glucosidase in Kluyveromyces fragilis. The enzyme biosynthesis was found to be non-adaptative, and repressed by glucose. These features are similar to those observed in K. fragilis. β-Glucosidase activity in the transformed yeast was much higher than in K. fragilis. We attempted to ferment cellobiose with the transformed yeast: practically no cellobiose was consumed, growth and ethanol production were negligible. Warburg experiments showed that cellobiose fermentation did not occur when the respiratory chain was not functioning.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00402336
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  • 6
    Electronic Resource
    Electronic Resource
    Killer toxin producing strains of the yeasts Hanseniaspora uvarum and Pichia kluyveri (1988)
    Springer
    Archives of microbiology 149 (1988), S. 261-267 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Hanseniaspora uvarum ; Pichia kluyveri ; Killer toxin ; dsRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By heat treatment killer strains of the type K1 of Saccharomyces cerevisiae that are known to harbour dsRNA plasmids were completely cured, whereas only a small fraction of the clones of the killer type K2 had lost the dsRNA dependent killer character. The K2 killers but not the strains of killer type K1 were easily cured by cycloheximide. Killer strains of Hanseniaspora uvarum were not curable by heat treatment. Curing was successfull with cycloheximide or 5-fluorouracil. Two double-stranded RNA plasmids were detected in the killer strains of H. uvarum. The smaller dsRNA plasmid was absent in the strains that were cured of their killer character by 5-fluorouracil. The killer character of H. uvarum was transferred to S. cerevisiae by spheroplast fusion. The fusion products showing the killer character contained both dsRNA plasmids, obviously the smaller plasmid (M-dsRNA) carries the genes for killer toxin formation. Killer strains of Pichia kluyveri were not curable of their killer character, in these strains no dsRNA plasmids were detected.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422015
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  • 7
    Electronic Resource
    Electronic Resource
    Immunological evidence for the involvement of cell wall proteins in phosphate uptake in the yeast Saccharomyces cerevisiae (1986)
    Springer
    Archives of microbiology 144 (1986), S. 207-212 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Phosphate uptake ; Antigenic relationships ; Cell wall proteins ; Candida tropicalis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunological cross-reactivity between cell wall proteins obtained from two yeast genera (Candida tropicalis and Saccharomyces cerevisiae) is reported. Specific retention of two cell wall proteins from Saccharomyces cerevisiae by an immunoabsorbent column coupled with antibodies against phosphate binding protein 2 (PiBP2) from Candida tropicalis allowed to generate antibodies against the proteins from S. cerevisiae. These antibodies were effective in inhibiting phosphate uptake by S. cerevisiae cells. The proteins from S. cerevisiae displayed a phosphate binding activity which was inhibited in the presence of the forementioned antibodies. These results and the observation that the amount of these proteins in the shock fluid was dependent of the growth conditions (i.e., in the presence or in the absence of phosphate) support the idea that these proteins are involved in the high affinity phosphate transport system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00410948
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  • 8
    Electronic Resource
    Electronic Resource
    Sensitivity of yeast glycolytic enzymes to chloroquine (1988)
    Springer
    Archives of microbiology 150 (1988), S. 309-312 
    Details
    ISSN: 1432-072X
    Keywords: Chloroquine ; Glycolytic enzymes ; Yeast ; Chloroquine and ATP/ADP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chloroquine at pH 8.0 and 10 mM concentration inhibits about 30% glucose consumption and ethanol formation in yeast cells. Out of the 11 glycolytic enzymes assayed, phosphoglycerate kinase and pyruvate decarboxylase have been found to be most sensitive to chloroquine. Next sensitive are hexokinase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase. Kinetic studies with the three kinases studied revealed competitive inhibition of chloroquine with ATP (hexokinase, phosphoglycerate kinase) or ADP (pyruvate kinase).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00407797
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  • 9
    Electronic Resource
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    A malic acid dependent mutant of Schizosaccharomyces malidevorans (1989)
    Springer
    Archives of microbiology 152 (1989), S. 564-566 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425487
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  • 10
    Electronic Resource
    Electronic Resource
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00415269
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  • 11
    Electronic Resource
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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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00446752
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  • 12
    Electronic Resource
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    How hexoses and inhibitors influence the malate transport system in Zygosaccharomyces bailii (1988)
    Springer
    Archives of microbiology 150 (1988), S. 37-41 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Hexose transport ; Sugar ; Malate uptake ; 2,4-DNP ; Zygosaccharomyces bailii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When grown in fructose or glucose the cells of Zygosaccharomyces bailii were physiologically different. Only the glucose grown cells (glucose cells) possessed an additional transport system for glucose and malate. Experiments with transport mutants had lead to the assumption that malate and glucose were transported by one carrier, but further experiments proved the existence of two separate carrier systems. Glucose was taken up by carriers with high and low affinity. Malate was only transported by an uptake system and it was not liberated by starved malate-loaded cells, probably due to the low affinity of the intracellular anion to the carrier. The uptake of malate was inhibited by fructose, glucose, mannose, and 2-DOG but not by non metabolisable analogues of glucose. The interference of malate transport by glucose, mannose or 2-DOG was prevented by 2,4-dinitrophenol, probably by inhibiting the sugar phosphorylation by hexokinase. Preincubation of glucose-cells with metabolisable hexoses promoted the subsequent malate transport in a sugar free environment. Preincubation of glucose-cells with 2-DOG, but not with 2-DOG/2,4-DNP, decreased the subsequent malate transport. The existence of two separate transport systems for glucose and malate was demonstrated with specific inhibitors: malate transport was inhibited by sodium fluoride and glucose transport by uranylnitrate. A model has been discussed that might explain the interference of hexoses with malate uptake in Z. bailii.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00409715
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  • 13
    Electronic Resource
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    Chloroquine, a lysosomotropic agent, inhibits zygote formation in yeast (1988)
    Springer
    Archives of microbiology 151 (1988), S. 20-25 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating ; Zygote formation ; Chloroquine ; Lysosomotropic agent ; Plasma membrane ; Cell fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Haploid cells of opposite mating type of Saccharomyces cerevisiae conjugate to form zygote. During the conjugation process, the degradation or reorganization of the cell wall and the fusion of the two plasma membranes take place. Since chloroquine inhibits cellular events associated with the reorganization of the plasma membrane, the effect of the drug on conjugation was studied. Chloroquine at a concentration, at which cell growth was not retarded, inhibited zygote formation, while it did not affect other mating functions, such as sexual agglutination, production of and response to mating pheromone. Cells in a mating culture containing chloroquine formed no “prezygote” suggesting that they were not prepared for entering into fusion process. The inhibitory effect of chloroquine was reversible as cells formed zygote when they were washed after treatment with chloroquine. Zygote formation was unaffected in cells possessing chlorquine within vacuoles after incubation with the drug in complete medium (YPD) at pH 7.5, followed by washing. This suggests that chloroquine inhibits zygote formaton by adsorbing to the plasma membrane of S. cerevisiae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00444663
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  • 14
    Electronic Resource
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    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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  • 15
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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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00456092
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  • 16
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    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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  • 17
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    The origin of mutant cells: mechanisms by whichSaccharomyces cerevisiae produces cells homoplasmic for new mitochondrial mutations (1985)
    Springer
    Current genetics 9 (1985), S. 627-640 
    Details
    ISSN: 1432-0983
    Keywords: Mitochondria ; Mutation ; Yeast ; Selection ; Random drift
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Haploid yeast cells have about 50 copies of the mitochondrial genome, and a mutational event is unlikely to affect more than one of these at a time. This raises the question of how such cells, or their progeny, become fixed (homoplasmic) for the mutant alele. We have tested the roles of six hypothetical mechanisms in producing erythromycin-resistant mutant cells: (i) random partitioning of mitochondrial genomes at cell division; (ii) intracellular selection for mtDNA molecules of one genotype; (iii) intracellular random drift of mitochondrial allele frequencies; (iv) intercellular selection for cells of a particular mitochondrial genotype; (v) induction of mitochondrial gene mutations by the antibiotic used to select mutants; and (vi) reduction in the number of mitochondrial genomes per cell by the antibiotic. Our experiments indicate that intracellular selection plays the major role in producing erythromycin-resistant mutant cells in the presence of the antibiotic. In the absence of the antibiotic, the combined effects of random drift and random partitioning are most important in determining the fate of new mutations, most of which are lost rather than fixed. Our experiments provide no evidence for mutation induction or ploidy reduction by erythromycin.
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    URL: http://dx.doi.org/10.1007/BF00449815
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  • 18
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    Efficient expression of theEscherichia coli leuB gene in yeast (1985)
    Springer
    Current genetics 9 (1985), S. 653-660 
    Details
    ISSN: 1432-0983
    Keywords: Gene expression ; Yeast ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Efficient expression of theEscherichia coli ZeuB (ß-isopropylmalate dehydrogenase) gene occured in yeast after in vitro DNase digestion and religation of plasmid bound ZeuB and the yeastIIIS3 DNA which placed the 5′ end of the yeastHIS3 gene immediately adjacent to the coding region of theE. coli leuB gene. Two structurally distinct classes of gene fusions were constructed, each involved portions of the yeastHIS3 gene which contributed DNA sequences responsible forleuB expression in yeast. The first class involved fusion of theHIS3 coding region to bacterial DNA resulting in the production of a fusion protein with ß-isopropylmalate dehydrogenase activity. The second class consisted of bacterial DNA, including theleuB coding region, fused to theHIS3 promotor region with the absence of any portion of theIIIS3 coding region. In both constructions theIIIS3 promotor region is required for transcription, however, translation of the class two fusion is initiated at a bacterial DNA coded AUG, and the 5′ end of the mRNA coded by theleuB gene mapped predominantly at bacterial DNA sequences. The DNA sequence responsible for the 5′ end of theHIS3 mRNAs remain in the class two gene fusions but this did not preclude the initiation of transcription at bacterial DNA sequences. The pattern of mRNA initiation at bacterial DNA suggests that DNA sequences at, or adjacent to, the site of transcription initiation are involved in the determination of the sites of initiation, and perhaps the frequency at which initiation occurs.
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    URL: http://dx.doi.org/10.1007/BF00449818
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  • 19
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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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569407
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  • 20
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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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569423
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  • 21
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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 
    Details
    ISSN: 1476-5535
    Keywords: Single cell protein ; Sucrose ; Yeast ; Thermotolerance ; Fermentation ; Kluyveromyces marxianus var.marxianus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Amino acid analyses were undertaken on single cell protein (SCP) produced by thermotolerant strains ofKluyveromyces marxianus var.marxianus grown on sugar cane molasses at 40°C. The maximum conversion of available sugars to biomass at 45°C was only 10.8% (g dry wt.·g−1 total sugars). The amino acid composition of the SCP did not differ markedly from that reported for other yeast species.
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    URL: http://dx.doi.org/10.1007/BF01569436
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  • 22
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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 
    Details
    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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    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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    Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation (1988)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
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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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    Cryptic DNA plasmids of the heterothallic yeast Saccharomycopsis crataegensis (1987)
    Springer
    Current genetics 12 (1987), S. 297-304 
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    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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    Phenotypic suppression and nuclear accommodation of the mit − oxi1-V25 mutation in isolated yeast mitochondria (1987)
    Springer
    Current genetics 12 (1987), S. 305-310 
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    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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    A new mutation for multiple drug resistance and modified plasma membrane ATPase activity in Schizosaccharomyces pombe (1986)
    Springer
    Current genetics 10 (1986), S. 359-364 
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    ISSN: 1432-0983
    Keywords: Multiple drug resistance ; ATPase ; Yeast ; Plasma membrane ; Cycloheximide ; pma ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mutant JV66 was selected from the wild type strain of S. pombe 972h− ade7-413 by its ability to grow on solid rich medium containing 200 μg Dio-9/ml. The single nuclear mutation, designated pma1 gives resistance towards diguanidines and several other positively charged compounds. The pma1 mutation also decreases plasma membrane ATPase activity and confers resistance of ATPase to vanadate. The pma1 locus is localized on chromose I at 5.3 map units from cyh1-C7 and at about 20.7 map units from the centromere. This new mutation is genetically and phenotypically different from the mutation cyh3 and cyh4 previously described (Johnston and Coddington 1983).
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    Genetic mapping of eleven spo genes essential for ascospore formation in the fission yeast Schizosaccharomyces pombe (1986)
    Springer
    Current genetics 10 (1986), S. 443-447 
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    ISSN: 1432-0983
    Keywords: Mapping ; Sporulation ; Yeast ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sporulation-deficient mutants of the fission yeast Schizosaccharomyces pombe were isolated from a homothallic strain mutagenized with ethyl methanesulfonate. Complementation tests defined two new genetic loci (spo19 and spo20) essential for ascospore formation, in addition to the 18 known spo loci (Bresch et al. 1968). A novel mapping procedure using random spore analysis prior to tetrad analysis allowed us to map 11 spo genes. Four genes (spo3, spo15, spo19 and spo20) were mapped on chromosome I, 6 genes (spo2, spo4, spoS, spo6, spo14 and spo18) on chromosome II and 1 gene (spo13) on chromosome III. Although there was no noticeable clustering of spo genes on the chromosomes, three pairs of linked genes (spo15-spo20, spo3-spo19 and spo2-spo18) were found.
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    Suppression of temperature sensitive mutations in oncogene-related CDC genes in Saccharomyces cerevisiae by catabolite repression resistance and cytoplasmic petite mutations (1985)
    Springer
    Current genetics 10 (1985), S. 35-37 
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    ISSN: 1432-0983
    Keywords: Yeast ; Carbon catabolite repression ; Oncogene-related genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The “start” cell division control genes CDC36 and CDC28 have been reported to contain a certain sequence homology to tissue oncogenes (ets and some protein kinase encoding oncogenes respectively). Here we report that temperature sensitive mutations in these genes are suppressed in cytoplasmic “petite” mutants and catabolite repression resistant mutants.
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    The cellular level of yeast ribosomal protein L25 is controlled principally by rapid degradation of excess protein (1986)
    Springer
    Current genetics 10 (1986), S. 733-739 
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    ISSN: 1432-0983
    Keywords: Yeast ; Ribosome synthesis ; Regulation ; Ribosomal protein turnover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When the gene dosage for the primary rRNA-binding ribosomal protein L25 in yeast cells was raised about 50-fold, the level of mature L25 transcripts was found to increase almost proportionally. The plasmid-derived L25 transcripts were structurally indistinguishable from their genomic counterparts, freely entered polysomes in vivo and were fully translatable in a heterologous in vitro system. Nevertheless, pulse-labelling for periods varying from 3–20 min did not reveal a significant elevation of the intracellular level of L25 protein. When pulse-times were decreased to 10–45 s, however, we did detect a substantial over production of L25. We conclude that, despite the strong RNA-binding capacity of the protein, accumulation of L25 is not controlled by an autogenous (pre-)mRNA-targeted mechanism similar to that operating in bacteria, but rather by extremely rapid degradation of excess protein produced.
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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 
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    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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    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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    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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    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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    Alkylation mutagenesis in Saccharomyces cerevisiae: lack of evidence for an adaptive response (1986)
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    Current genetics 10 (1986), S. 647-655 
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    ISSN: 1432-0983
    Keywords: Alkylation mutagenesis ; Adaptive response ; rad6 ; rad52 ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have found no evidence for an adaptive response for either lethality or mutagenesis following treatment of Saccharomyces cerevisiae with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). The rad6 and rad52 mutants of S. cerevisiae are highly defective in MNNG and ethyl methanesulfonate induced mutagenesis of both stationary and exponential phase cells. These and other observations indicate that the mechanisms of repair of alkylation damage and mutagenesis differ markedly between S. cerevisiae and Escherichia coli.
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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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    The Saccharomyces cerevisiae RAD2 gene complements a Schizosaccharomyces pombe repair mutation (1989)
    Springer
    Current genetics 15 (1989), S. 27-30 
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    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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    Two nuclearly inherited loci conferring increased diuron resistance to NADH oxidase in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 31-38 
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    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.
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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 
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    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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    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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    An α-specific gene, SAG1 is required for sexual agglutination in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 393-398 
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    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.
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    Meiotic segregation of circular plasmid-minichromosomes from intact chromosomes in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 385-392 
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    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.
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    A novel class of vector for yeast transformation (1989)
    Springer
    Current genetics 16 (1989), S. 21-25 
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    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.
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    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.
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    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.
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    Structure of a linear plasmid of the yeast Kluyveromyces lactis; Compact organization of the killer genome (1985)
    Springer
    Current genetics 9 (1985), S. 147-155 
    Details
    ISSN: 1432-0983
    Keywords: Killer ; Yeast ; Linear plasmid ; Sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Some strains of the yeast Kluyveromyces lactis contain a pair of linear DNA plasmids, k1 and k2, 8.8 and 13.8 kilobase pairs long, respectively. Simultaneous presence of the two plasmids confer a killer phenotype on the cell by producing a toxin which blocks the growth of sensitive yeast species. Previous genetic studies have suggested that the toxin protein is coded by the k1 plasmid. We have now determined the total nucleotide sequence of k1 DNA. The genome is 8,874 base pairs in length. It contains four protein-coding reading frames, three transcribed from one strand and the fourth transcribed from the complementary strand and has terminal inverted repeats of 202 base pairs. Nuclease S1 mapping confirmed this arrangement and showed that these genes are transcribed. The terminal repeats and the four genes form an extremely compact genome, with some overlapping of genes. All four genes use highly biased codons, 86% of them having A or T at the wobble position, reminiscent of yeast mitochondrial genes. Three genes share a very similar 5′ leader sequence. The nature of gene products is discussed in the light of what is known of the excreted toxin protein.
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    Characterization of blasticidin S — resistant mutants of Saccharomyces cerevisiae (1985)
    Springer
    Current genetics 9 (1985), S. 179-181 
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    ISSN: 1432-0983
    Keywords: Blasticidin S ; Yeast ; Resistant mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Blasticidin S-resistant mutants of S. cerevisiae were isolated and characterized. Resistant mutations were found to fall into two complementation groups. A single recessive nuclear gene was responsible for each group, donated as bls1 and bls2, respectively. A gene bls1 was linked to an ilv3 gene located on the right arm of chromosome X. The resistant phenotypes from both genes were not associated with ribosomes known to be target sites of Blasticidin S, when analyzed by poly(U)-directed polyphenylalanine synthesis. The resistant mechanisms of the mutations are discussed in this paper.
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    The DEL1 mutator gene in Saccharomyces cerevisiae does not act in trans (1985)
    Springer
    Current genetics 9 (1985), S. 259-262 
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    ISSN: 1432-0983
    Keywords: Yeast ; Ty1 ; Trans ; Deletion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DEL1 strains of the yeast Saccharomyces cerevisiae exhibit a high rate of deletions of the three linked genes, CYC1, OSM1, and RAD7. Classical genetic methods showed that DELI segregated as a single Mendelian gene closely linked to CYC1. In addition, genetic evidence suggested that DEL1 was both cis- and trans-dominant (Liebman et al. 1979). Molecular analysis of deletions isolated from a haploid DEL1 strain established that deletion formation was mediated by recombination between yeast transposable elements, Ty's (Liebman et al. 1981). We now report the molecular characterization of deletions isolated from diploids in the trans configuration. This analysis reveals that these deletions probably arose in a two-step process involving mitotic recombination followed by Ty-mediated deletion formation in cis.
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    Virus-like particles from the yeast Yarrowia lipolytica (1985)
    Springer
    Current genetics 9 (1985), S. 279-284 
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    ISSN: 1432-0983
    Keywords: Virus-like particles ; Double-stranded RNA ; Yeast ; Yarrowia lipolytica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four out of the 24 strains of the yeast Yarrowia lipolytica we have checked for the presence of virus-like particles (VLPs) proved to contain encapsidated double-stranded RNA (dsRNA) molecules, 4.9 kb long. A major VLP polypeptide of MW 80,000 was observed in all 4 cases, and a second one of MW 77,000 in three cases. dsRNA from the VLPs harboring only the larger polypeptide showed little homology with the 3 others. We have found no homology between VLP dsRNAs and host DNA or dsRNAs from Saccharomyces cerevisiae, and no relationship between the presence of VLPs and a possible killer phenomenon in Y. lipolytica.
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    Conserved sequences upstream of yeast ribosomal protein genes (1985)
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    Current genetics 9 (1985), S. 273-277 
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    ISSN: 1432-0983
    Keywords: Yeast ; Ribosomal protein gene ; Sequence analysis ; Conserved elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Computer analysis has previously revealed the presence of a 12-nucleotide common sequence element (AACATC CA TG T A G CA; HOMOL1) in the upstream regions of several yeast ribosomal protein genes. By extending the sequence analysis of the 5′-flanking regions of a number of other ribosomal protein genes (including those encoding S10-1, S10-2, S33 and L16-2) we could establish that HOMOL1 occurs upstream of most but not all yeast ribosomal protein genes. Apart from HOMOL1 an additional conserved sequence (ACCCATACATT A T ; RPG-box) was detected in front of nearly all yeast ribosomal protein genes, although in some cases it is present in the opposite orientation in the other strand. There seems to be no correlation between the occurrence of one box and that of the other. However when both boxes are present the RPG-box is always located 3′ to the HOMOL1-sequence mostly at a distance of only a few nucleotides. A further one-to-one comparison of the upstream regions of several yeast ribosomal protein genes revealed extensive additional sequence homologies that are suggested to be involved in the coordinate control of ribosomal protein gene expression in yeast.
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    Selection and stability of yeast transformants expressing cDNA of an Ml killer toxin-immunity gene (1985)
    Springer
    Current genetics 9 (1985), S. 285-291 
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    ISSN: 1432-0983
    Keywords: Killer toxin ; Plasmid selection ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transformants of sensitive yeast strains containing an expressed cDNA copy of the yeast killer toxin-immunity gene could be selected for by exposure to added killer toxin. For strain AH-22 the transformation frequency was approximately 10% that obtained by selection for leucine prototrophy. The procedure required time for expression of immunity prior to selection, and a screening step to remove non-transformed survivors. Under conditions where active toxin was produced, transformants containing the toxin-immunity gene were at a selective advantage, and cells losing the plasmid were killed. This resulted in self selection of transformants, and affords a way of maintaining plasmid stability in protrophic strains.
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    Exposure of cdc8 homozygous diploids of Saccharomyces cerevisiae to nonpermissive temperature leads to an increase in mitotic conversion frequencies (1985)
    Springer
    Current genetics 9 (1985), S. 447-452 
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    ISSN: 1432-0983
    Keywords: Yeast ; cdc8-1 mutation ; Mitotic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In a diploid strain homozygous for the cdc8-1 mutation, a block in DNA synthesis caused by restrictive temperature resulted in a significant increase in the frequency of intragenic recombination at the HOM2 locus. Under restrictive conditions, incorporation of radioactivity into DNA was reduced to 2% of the control and alkaline sucrose gradient centrifugation revealed that only short DNA fragments were synthesized. There was no considerable fragmentation of template DNA during incubation of cdc8-1 strains under restrictive conditions.
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    Formation and stability of linear plasmids in a recombination deficient strain of yeast (1985)
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    Current genetics 9 (1985), S. 441-445 
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    ISSN: 1432-0983
    Keywords: Telomeres ; Recombination ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Natural termini from macronuclear DNA of the ciliated protozoans Tetrahymena thermophila and Oxytricha fallax can support telomere formation in yeast. However, plasmids carrying these ciliate termini are modified by the addition of DNA which hybridizes to the synthetic oligonucleotide poly [d(C-A)], a sequence which also hybridizes to terminal restriction fragments from yeast chromosomes but not to Tetrahymena or Oxytricha macronuclear DNAs. Thus, in yeast, the creation of new telomeres on ciliate termini involves the acquisition of yeast-specific terminal sequences presumably by either recombination or non-templated DNA synthesis. The RAD52 gene is required for the majority of yeast mitotic and meiotic recombination events. Moreover, the absence of an active RAD52 gene product results in high rates of chromosome loss. Here we demonstrate that terminal restriction fragments from Tetrahymena macronuclear ribosomal DNA (rDNA) support the formation of modified telomeres in a yeast strain carrying a defect in the RAD52 gene. Moreover, linear plasmids bearing these modified ciliate termini are stably propagated in rad52 − cells.
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    Either one of the two yeast EF-1α genes is required for cell viability (1985)
    Springer
    Current genetics 9 (1985), S. 693-697 
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    ISSN: 1432-0983
    Keywords: Yeast ; TEF genes ; Gene disruption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two genes,TEF1 andTEF2, encode the protein elongation factor EF-1α in the yeastSaccharomyces cerevisiae. We have generated yeast haploid strains containing eitherTEF1 orTEF2 interrupted by insertion of a large piece of foreign DNA. Cells which contain either one functional copy of the EF-1α genes are viable. In contrast, attempts to isolate a yeast haploid strain with bothTEF1 andTEF2 inactivated have failed suggesting that the double gene disruption is a lethal event.
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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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    Functional nuclear suppressor of mitochondrialoxi2 mutations in yeast (1985)
    Springer
    Current genetics 10 (1985), S. 87-93 
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    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; oxi2 mutations ; Functional suppressors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A semidominant nuclear suppressor, callednam6, ofoxi2-V276 mitochondrial mutation has been isolated and characterized. The nuclear character ofnam6 was proved by its retention inrho° strains, lack of mitotic segregation in diploids and meiotic 2:2 segregation in tetrads. The specificity ofnam6 was tested on 315mit − mutations of four mitochondrial genes (oxi1, oxi2, oxi3, andcob-box). It suppresses clearly only three mutations in theoxi2 gene, restoring partially or completely cytochrome aa3 formation. The results suggest a functional character of the suppression.
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    Transformation of ψ − Saccharomyces cerevisiae to ψ + with DNA co-purified with 3 μm circles (1986)
    Springer
    Current genetics 11 (1986), S. 79-82 
    Details
    ISSN: 1432-0983
    Keywords: Psi-factor ; 3-micron plasmids ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA enriched for supercoiled plasmids prepared from the 3 μm plasmid-enriched, [ψ +], [2 μm°] strain 6-1G-P188 and from the [2 μm+] [ψ+] strain LL20 can be used to transform a ψ − recipient strain to ψ +. Fractionation of the former preparation by electrophoresis showed that the 3 Mm plasmid band contained the transforming activity.
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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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    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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    Sporulation-regulated genes ofSaccharomyces cerevisiae (1985)
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    Current genetics 10 (1985), S. 163-169 
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    ISSN: 1432-0983
    Keywords: Sporulation ; Yeast ; Transcription ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have characterized 46 hybrid phage which hybridize preferentially to mRNA from sporulating cells. Cross-hybridization experiments demonstrate that 27 distinct SPR (Sporulation regulated) sequences are represented among these phage. The SPR genes can be grouped into three classes: early, middle, and late. The early class shows an accumulation of transcripts soon after transfer to sporulation medium and continues to accumulate RNA throughout sporulation. Transcripts of the middle class increase in level at about the time of DNA synthesis, rise rapidly in abundance until meiosis II, then accumulate more slowly for at least the next 3 h. Late gene transcripts begin to accumulate at about the time of meiosis I, increase 10- to 20-fold in the next 2 h, then remain constant in late sporulating cells.
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    Cloning and characterisation of the ribosomal RNA genes of the dimorphic yeast, Yarrowia lipolytica (1986)
    Springer
    Current genetics 10 (1986), S. 449-452 
    Details
    ISSN: 1432-0983
    Keywords: rRNA genes ; Yeast ; Yarrowia lipolytica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ribosomal RNA genes of Yarrowia lipolytica have been identified, both in restriction digests of total genomic DNA and in a pBR322 gene bank, by hybridisation with cloned Saccharomyces cerevisiae rDNA. The Y. lipolytica rDNA repeat unit is 8.9 kb in size and contains the genes for the 25S and 18S, but not the 5S, rRNA species. The number of copies of these repeat units is approx. 50 per haploid genome. Several clones were found which did not conform to the standard restriction map due to differences outside the coding region. It appears that there is either heterogeneity of the spacer sequence within a strain or that the Y. lipolytica rDNA genes may be present as a number of separate clusters within this yeast's genome.
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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 
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    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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    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.
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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 
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    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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    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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    URL: http://dx.doi.org/10.1007/BF00435458
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    Yeast arginine permease: nucleotide sequence of the CAN1 gene (1986)
    Springer
    Current genetics 10 (1986), S. 587-592 
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    ISSN: 1432-0983
    Keywords: Yeast ; Arginine permease ; Membrane protein ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast CAN1 gene, thought to encode arginine permease, has found use in genetics as a selectable locus. We have sequenced the cloned CAN1 gene, which contains an open reading frame of 1770 nucleotides, encoding a polypeptide of calculated molecular weight 65,766. Disruption of this open reading frame largely abolishes CAN1 gene expression, while subcloned fragments of the open reading frame hybridize strand —specifically to a 2.3 kb yeast RNA message. The encoded protein has no leader signal sequence, and is highly hydrophobic, with a possible twelve membrane-spanning domains, several of which have the high hydrophobic moments seen in channel-forming or permease proteins. This protein structure is consistent with the CAN1 product being the plasma membrane arginine permease.
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    URL: http://dx.doi.org/10.1007/BF00418125
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    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.
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    URL: http://dx.doi.org/10.1007/BF00447038
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    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.
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    URL: http://dx.doi.org/10.1007/BF00340710
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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 
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    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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    The influence of cell ploidy on the thermotolerance of Saccharomyces cerevisiae (1987)
    Springer
    Current genetics 11 (1987), S. 595-598 
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    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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    The gene for yeast ribosomal protein S31 contains an intron in the leader sequence (1985)
    Springer
    Current genetics 10 (1985), S. 1-5 
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    ISSN: 1432-0983
    Keywords: Posttranslational processing ; Ribosomal protein gene ; Transcript mapping ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Analysis of the primary structure of the gene for yeast ribosomal protein S31 revealed two unusual features. First, an intron of 312 nucleotides is located within the 5′-untranslated region. Second, the coding sequence for the known amino-terminal peptide of the protein starts 13 codons downstream of the ATG initiation codon, suggesting that S31 is synthesized as a precursor which undergoes post-translational processing to the mature protein. Primer extension analysis showed that transcription of the S31 gene starts at multiple sites. The 5′-flanking region of the gene contains several, previously described, conserved sequence elements that may play a role in the coordinate expression of yeast ribosomal protein genes.
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    URL: http://dx.doi.org/10.1007/BF00418486
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    Coincident chromosomal disomy in meiotic dyads from triploid yeast (1987)
    Springer
    Current genetics 12 (1987), S. 569-576 
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    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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    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 
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    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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    Identification of the yeast DNA polymerase I gene with antibody probes (1985)
    Springer
    Current genetics 10 (1985), S. 245-252 
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    ISSN: 1432-0983
    Keywords: DNA polymerase ; Yeast ; Immunoscreening ; Cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Partially overlapping fragments of the gene encoding yeast DNA polymerase I have been cloned by immunological screening of a yeast genomic library constructed in the phage λ expression vector λgt11. The three gene fragments we analyzed in detail encode part of a yeast protein that has been identified as yeast DNA polymerase I, because it shares with this enzyme a number of antigenic determinants. In fact, the yeast protein fragments expressed by the recombinant phages react with both polyclonal and monoclonal antibodies raised against different, highly purified preparations of DNA polymerase I. Moreover, they can be used to affinity purify antibodies specifically reacting with active DNA polymerase I polypeptides and they compete with the yeast enzyme for binding to antibodies that inhibit catalytic activity. The gene is located on chromosome XIV in the yeast genome, and it is transcribed as a 5.2 kb mRNA.
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    Deletion mapping of the yeast pol I promoter (1985)
    Springer
    Current genetics 10 (1985), S. 253-260 
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    ISSN: 1432-0983
    Keywords: Yeast ; RNA polymerase I ; Promoter ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Deletions in the promoter region of the 37S pre-rRNA operon in yeast were constructed and analysed in vivo using an artificial ribosomal minigene present on an extrachromosomal yeast vector. Sequences required for correct transcription initiation were found to be located between positions −192 and +15 relative to the start; a 5′-deletion down to position −133 reduces the transcription yield of the minigene at least five-fold. To allow detection of transcription of the minigene in isolated nuclei of yeast transformed with a minigene-bearing plasmid we attempted to increase the minigene copy number. The transcription yield in vivo appeared not to be proportional to the copy number but was found to be greatly enhanced when two or three mini-genes are present in tandem. α-Amanitin sensitivity of transcription of these minigenes in isolated nuclei proved that RNA polymerase I is responsible for their transcription.
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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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    FLP recombinase induction of the breakage-fusion-bridge cycle and gene conversion in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 273-281 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; FLP-FRT ; BFBC ; Gene conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A YEp chimaeric plasmid containing URA3 and SMR1 [sulfometuron methyl resistant (SMR) allele of ILV2] as selectable markers, and the 2 μm site-specific recombination FLP recognition target (FRT), was integrated at the ilv2-Δ1 site in chromosome XIII in a cir°] haploid. Southern analysis defined two integrant structures. Structure I had URA3 distal and SMR1 proximal to FRT whereas in structure II both markers were distal to FRT. Selectable markers were stably inherited in [cir°] haploids and [cir°] diploids heterozygous for the integrant and ILV2. Approximately 14% of heterozygous [cir +] diploid cells exhibited homozygotization for the distal (500 kb) ade4 marker in trans. In [cir +] diploids FLP-FRT recombination resulted in the simultaneous loss of both structure II markers, whereas the structure I distal URA3 marker loss always preceded the variable loss of the proximal SMR1 marker. URA− cells continued to segregate for loss of SMR1 until stable URA− SMR or URA−SMS cells were produced. Gene conversion was identified in stable URA−SMR cells that were homozygous SMR1/SMR1 but contained wild type ILV2 restriction endonuclease sites. These observations support a model based on concerted FLP-FRT action resulting from the secondary integration of native 2 μm DNA followed by unequal sister chromatid exchange (USCE) within inverted FRTs. The resultant chromatid bridge resulted in a double-stand break. Fusion of the broken ends of sister chromatids generated a breakage-fusion-bridge cycle (BFBC). Repeated rounds of the BFBC resulted in proximal marker loss and the generation of additional double-strand breaks. Recombinogenic properties of the double-strand break initiated events leading to homozygotization and gene conversion.
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    Comparison of expression of the endo-β-1,3-1,4-glucanase gene from Bacillus subtilis in Saccharomyces cerevisiae from the CYC1 and ADH1 promoters (1986)
    Springer
    Current genetics 11 (1986), S. 65-70 
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    ISSN: 1432-0983
    Keywords: Yeast ; β-glucanase ; Bacillus ; Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The endo-β-1,3-1,4-glucanase gene from B. subtilis was placed under yeast promoter control in a number of different yeast expression vectors. The hybrid plasmids were transformed into S. cerevisiae where they directed the synthesis of varying amounts of active enzyme. The presence of B. subtilis DNA sequences 5′ to the initiation codon for the B. subtilis β-glucanase gene reduced expression of the gene in yeast. A 1,000-fold increase in the yield of β-glucanase was obtained using the ADH1 promoter compared with the CYC1 promoter.
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    URL: http://dx.doi.org/10.1007/BF00389427
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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 
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    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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    URL: http://dx.doi.org/10.1007/BF00378177
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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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    Cloning of a nuclear gene MRS1 involved in the excision of a single group I intron (bI3) from the mitochondrial COB transcript in S. cerevisiae (1986)
    Springer
    Current genetics 11 (1986), S. 185-191 
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    ISSN: 1432-0983
    Keywords: Gene cloning ; Mitochondrial RNA splicing ; Nuclear mutants ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The respiratory deficient yeast nuclear mutant MK3 is defective in the synthesis of the mature transcripts of the mitochondrial COB and OX13 genes, which code for apocytochrome b and subunit I of cytochrome c oxidase, resp. Introns 3 and 4 of the COB transcript (bI3 and bI4) and intron 4 (aI4) of the OXI3 transcript can not be excised (Pillar et al. 1983a, b). When combined with mitochondrial genomes lacking introns bI1, bI2 and bI3, or lacking intron bI3 alone the mutant is respiratory competent. Thus, the non-excision of bI4 and aI4 turns out to be an indirect effect of the mutation. From a wild type yeast genebank a plasmid has been isolated with a 3.3 kb DNA insert, which complements the mutant. Subcloning experiments assigned the functional gene to a 1.6 kb HaeIII-Sau3A fragment. Hybridization experiments showed, that it is (i) a single copy gene, (ii) also present in strain D273-10B, containing the “short form” mitochondrial genome (lacking the COB introns bI1-bI3), and (iii) located on chromosome IX. The nuclear gene defective in mutant MK3, was named MRS1 (Mitochondrial RNA Splicing). The involvement of this nuclear gene in the excision of a single group I mitochondrial intron (bI3) of the COB transcript is discussed.
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    DNA repair genes of Saccharomyces cerevisiae: complementing rad4 and rev2 mutations by plasmids which cannot be propagated in Escherichia coli (1986)
    Springer
    Current genetics 11 (1986), S. 205-210 
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    ISSN: 1432-0983
    Keywords: Yeast ; Excision repair ; Mutagenic DNA repair ; RAD4 and REV2 gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The RAD4 gene of yeast required for the incision step of DNA excision repair and the REV2 (= RAD5) gene involved in mutagenic DNA repair could not be isolated from genomic libraries propagated in E. coli regardless of copy number of the shuttle vector in yeast. Transformants with plasmids conferring UV resistance to a rad4-4 or a rev2-1 mutant were only recovered if yeast was transformed directly without previous amplification of the gene bank in E. coli. DNA preparations from these yeast clones yielded no transformants in E. coli but retransformation of yeast was possible. This lead to the isolation of a defective derivative of the rad4 complementing plasmid. The modified plasmid was now capable of transforming E. coli but still interfered significantly with its growth.
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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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    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 
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    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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    Mutational analysis of the upstream activation site of yeast ribosomal protein genes (1989)
    Springer
    Current genetics 15 (1989), S. 247-251 
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    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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    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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    URL: http://dx.doi.org/10.1007/BF02427750
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    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.
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    URL: http://dx.doi.org/10.1007/BF00435502
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  • 89
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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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  • 90
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    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 
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    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.
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    URL: http://dx.doi.org/10.1007/BF00340714
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  • 91
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    High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier (1989)
    Springer
    Current genetics 16 (1989), S. 339-346 
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    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.
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    URL: http://dx.doi.org/10.1007/BF00340712
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  • 92
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    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 
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    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.
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    URL: http://dx.doi.org/10.1007/BF00340711
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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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    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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    Cloning of the orotidine 5′-phosphate decarboxylase (ODC) gene of Schwanniomyces occidentalis by complementation of the ura3 mutation in S. cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 29-35 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Cloning ; ODC ; Complementation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A DNA fragment containing the gene encoding orotidine 5′-phosphate decarboxylase (ODC) from the yeast, Schwanniomyces occidentalis (formerly castellii) has been isolated from a genomic library constructed in the S. cerevisiae expression vector, pYcDE8. A recombinant plasmid, p2-lA, containing a 2.47 kb insert was shown to complement the ura3-52 mutation of several strains of S. cerevisiae. This DNA insert was shown to be from Schwanniomyces occidentalis by Southern hybridization analysis. A restriction enzyme cleavage map of the insert has been derived and the ODC gene localized to a 1.1 kb region by deletion analysis. In addition, we have demonstrated that expression of ODC is not dependent on the ADHI promoter carried on pYcDE8. This is the first report of the cloning of a gene from a member of the genus Schwanniomyces.
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    URL: http://dx.doi.org/10.1007/BF00365753
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    Allelism between pso1-1 and rev3-1 mutants and between pso2-1 and snm1 mutants in Saccharomyces cerevisiaee (1988)
    Springer
    Current genetics 13 (1988), S. 37-40 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; DNA repair mutants ; Allelism test ; Psoralen plus UVA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the yeast Saccharomyces cerevisiae, allelism between the psol-1 and the rev3-1 mutants on the one hand and the pso2-1 and snm1 mutants on the other, is demonstrated by the comparison of phenotypes, complementation tests and meiotic segregation analysis.
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    Deletions in the cob gene of yeast mtDNA and their phenotypic effect (1985)
    Springer
    Current genetics 10 (1985), S. 283-290 
    Details
    ISSN: 1432-0983
    Keywords: Mitochondria ; Yeast ; Deletions ; RNA stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two cob − deletion mutants are characterized. One of them, M9410, is deleted for 911 by of the noncoding sequences only which separate tRNAGlu and cob exon 1; it thus lacks most of the sequence encoding the 957 by long cob leader (Bonitz et al. 1982) and some 20 by 5′ to it. The end points of this deletion coincide with 31 by long direct repeats in wild type mtDNA. The other mutant, M9391, is deleted for all cob coding sequences and most of the cob leader sequence but it retains the 5′ terminal 261 by of this leader. Northern analysis revealed that M9410 totally lacks cob mRNA or pre-mRNA. The large deletion M9391 in contrast accumulates a 13S RNA which probably results from transcription through the junction, which ligates sequences of the cob leader to sequences of the cob-oli1 intergenic spacer.
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    URL: http://dx.doi.org/10.1007/BF00365624
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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 
    Details
    ISSN: 1432-0983
    Keywords: Ribosomal protein ; Immunological homology ; Yeast ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Polyclonal antibodies raised against ribosomal protein (r-protein) L2 of Schizosaccharomyces pombe were used to check for cross-reaktions with total r-proteins of rat liver. Using this procedure, the rat liver r-proteins, L4 and L24, were identified as being immunologically related to yeast L2. In addtional, homologies between rat liver L4 and L24 were detected. The possible implications for the regulation of r-protein synthesis are discussed.
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    URL: http://dx.doi.org/10.1007/BF00387769
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  • 99
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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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  • 100
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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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    URL: http://dx.doi.org/10.1007/BF00355403
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