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  • Saccharomyces cerevisiae  (87)
  • Springer  (87)
  • Cell Press
  • 1985-1989  (72)
  • 1980-1984  (15)
  • 1988  (38)
  • 1986  (34)
  • 1983  (15)
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  • 1985-1989  (72)
  • 1980-1984  (15)
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  • 1
    Electronic Resource
    Electronic Resource
    High frequency FLP-independent homologous DNA recombination of 2 μ plasmid in the yeast Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 191-199 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Site specific recombination ; 2 μ DNA plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The purpose of this work is to identify and quantitate in vivo 2 μ plasmid FLP-independent recombination in yeast, using a nonselective assay system for rapid detection of phenotypic expression of the recombination events. A tester plasmid was constructed such that in vivo recombination between 2 μ direct repeat sequences produces the resolution of the plasmid into two circular DNA molecules. This recombinational event is detected as a phenotypic shift from red to white colonies, due to the mitotic loss of the plasmid portion containing the yeast ADE8 gene in a recipient ade1 ade2 ade8 genetic background. In the absence of the 2 μ FLP recombinase and/or its target DNA sequence, recombination is not abolished but rather continues at a high frequency of about 17%. This suggests that the FLP-independent events are mediated by the chromosomally-encoded general homologous recombination system. We therefore conclude that the totality of 2 μ DNA recombination events occurring in FLP+ cells is the contribution of both FLP-mediated and FLP-independent events.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00376739
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  • 2
    Electronic Resource
    Electronic Resource
    Molecular cloning of the ARG7 gene of Schizosaccharomyces pombe encoding argininosuccinate lyase (1988)
    Springer
    Current genetics 14 (1988), S. 381-385 
    Details
    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Argininosuccinate lyase ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A gene bank of Sau3A partially restricted Schizosaccharomyces pombe DNA in YEp13 was used to transform an arg4 mutant of Saccharomyces cerevisiae. One colony was recovered which contained the YEp13 plasmid bearing a large insert complementing the argininosuccinate lyase (ASL) mutation. As shown by restriction mapping and subcloning experiments, the DNA sequence required for complementation is localized on a 2 kb BamHI-BamHI fragment. The plasmid complemented several S. cerevisiae arg4 mutants of independent origin and a S. pombe arg7 mutant lacking ASL. Low but significant ASL activities were detected in crude extracts of these transformants. No complementation of the E. coli argH mutant was observed. Southern blot hybridizations showed that the insert originates from the S. pombe genome. No cross-hybridization was found between this sequence and S. cerevisiae DNA. It can be concluded that the cloned DNA fragment bears the S. pombe ARG7 gene coding for ASL.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419996
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  • 3
    Electronic Resource
    Electronic Resource
    Inheritance of chromosome length polymorphisms in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 413-418 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome length polymorphisms ; FIGE ; OFAGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Although Saccharomyces cerevisiae strains generally have similar chromosomal band patterns as revealed by pulsed field gel electrophoresis, individual bands often move slightly differently from one strain to the other. Surveying strains from our stock collection, we found that nearly all the bands of a certain pair of strains differed in their mobility. Some of these chromosome length polymorphisms segregated in a 2:2 ratio, indicating that they resulted from single structural alterations (i.e. additions or deletions). One of these was mapped on the right arm of chromosome 1. Others did not segrate in a simple 2:2 ratio. That is, there were progenies which had bands not present in either parent. We suggest that these new bands are the products of recombination between homologous chromosomes having two or more structural alterations.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00521262
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  • 4
    Electronic Resource
    Electronic Resource
    Comparison of Tetrahymena ARS sequence function in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe (1988)
    Springer
    Current genetics 14 (1988), S. 225-233 
    Details
    ISSN: 1432-0983
    Keywords: ARS ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Tetrahymena thermophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated several Tetrahymena thermophila chromosomal DNA fragments which function as autonomously replicating sequences (ARS) in the heterologous Saccharomyces cerevisiae and Schizosaccharomyces pombe selection systems. The Tetrahymena ARS sequences were first isolated in S. cerevisiae and were derived from non-ribosomal micro- and macronuclear DNA. Sequence analysis of the ARS elements identified either perfect or close matches with the 11 by S. cerevisiae ARS core consensus sequence. Subcloning studies of two Tetrahymena ARS elements defined functional regions ranging in size from 50 to 300 bp. Testing of the ARS elements in S. pombe revealed that most of the T. thermophila inserts confer ARS function in both yeasts, at least in the sense of promoting a high transformation frequency to plasmids which contain them. However, the actual sequences responsible for ARS activity were not always identical in the two yeasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00376742
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  • 5
    Electronic Resource
    Electronic Resource
    Construction and characterization of a haploid strain of Saccharomyces cerevisiae that completely lacks all genomic CYH2 sequences (1988)
    Springer
    Current genetics 14 (1988), S. 325-329 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Ribosomal protein genes ; CYH2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A diploid strain of the yeast Saccharomyces cerevisiae has been constructed that has one copy of the ribosomal protein gene CYH2 completely deleted and replaced with the TRP1 gene using the method of Rothstein (1983). There are only small differences in growth rate and no detectable difference in steady state level of CYH2 mRNA between the diploid that is heterozygous for the CYH2 deletion and the parent diploid with two normal copies of this gene. This suggests that the diploid must partially compensate for the loss of one CYH2 gene. Tetrad dissection shows that haploid spores lacking the CYH2 gene cannot germinate. The lethality of this deletion can be rescued by a CYH2 cDNA on a low copy vector. Haploids which lack the genomic copy of the CYH2 gene, but contain a plasmid copy of the CYH2 cDNA are able to grow normally. These CYH2 deleted yeast haploids should be useful to analyze mutationally altered CYH2 genes and genes homologous to CYH2 from other organisms without interference from a genomic copy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419989
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  • 6
    Electronic Resource
    Electronic Resource
    The glucose- and ethanol-dependent regulation of PDC1 from Saccharomyces cerevisiae are controlled by two distinct promoter regions (1988)
    Springer
    Current genetics 14 (1988), S. 337-344 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Gene regulation ; Saccharomyces cerevisiae ; PDCI promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 870 by promoter fragment of the PDC1 gene that includes the carbon source dependent regulatory regions was investigated using 5′ and 3′ promoter deletions. The results indicate that glucose and ethanol regulation of PDC1 transcription are independently controlled by distinct cis-acting regions. The consensus sequence AAATCGATA may play a role in this regulation, while the sequence (ATCA)AACCT may be important in transcription initiation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419991
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  • 7
    Electronic Resource
    Electronic Resource
    A colony procedure for transformation of Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 21-23 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Transformation ; Plasmid ; Colony ; Polyethylene glycol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid and simple yeast transformation procedure has been developed using colonies on agar plates. Saccharomyces cerevisiae SHY3 cells were picked up from colonies on YPD plates grown freshly or stored at 4 °C and incubated with M13RK9-T DNA at 30 °C for 1–2 h in a solution of Li+, Ca2+, Mg2+, triacetin and polyethylene glycol. About 3,500 transformants were obtained per µg of double stranded M13RK9-T DNA. Unlike the existing spheroplast techniques, single stranded M13RK9-T DNA transformed intact cells below one-hundredth frequency of the duplex form.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365751
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  • 8
    Electronic Resource
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    Cosmids carrying Aspergillus terreus DNA can integrate into Saccharomyces cerevisiae chromosome XII via recombination between yeast and foreign DNAs (1988)
    Springer
    Current genetics 14 (1988), S. 183-189 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus terreus clonotheque ; Saccharomyces cerevisiae ; Homologous integration ; 2 μ circledirected chromosome destabilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A genome clonotheque consisting of 25- to 40-kb Sau3A1 fragments of Aspergillus terreus DNA was constructed in the episomal cosmid vector pES33 containing the yeastARG4 gene. From the 475 transformants of cir° yeast strain ESH-0, 23 stable Arg + transformants were independently selected. Genetic and Southern analysis of these stable transformants showed that 39% arose as a result of recombination between cloned A. terreus DNA sequences and yeast chromosome XII. The recombination events most likely occurred in the regions of homology within the rDNA clusters of A. terreus and Saccharomyces cerevisiae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00376738
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  • 9
    Electronic Resource
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    Generation of an ilv bradytrophic phenocopy in yeast by antisense RNA (1988)
    Springer
    Current genetics 13 (1988), S. 283-289 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Inducible antisense gene ; Acetolactate synthase ; Bradytrophic phenocopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report for the first time on the regulation of gene expression in yeast by antisense RNA. Chimaeric genes were constructed containing the 5′ upstream and partial coding sequence of SMR1 — a sulfometuron methyl resistant allele of the ILV2 locus. Such fragments were placed 5′ to 3′ and 3′ to 5′ under control of the GAL10 promoter and CYCl terminator in a high copy YEp plasmid. Following galactose induction only transformants containing antisense RNA genes showed biological activity against SMR1 gene expression. Antisense RNA inhibited synthesis of the SMR1 gene product acetolactate synthase and thus repressed cellular growth which resulted in a bradytrophic auxotroph revertable by addition of isoleucine and valine. Antisense RNA inhibition was enhanced in galactose medium containing sulfometuron methyl and in gcn4 cells deficient for positive regulation of the ILV2 locus. This system can be used to study factors that interfere with antisense RNA function and to assign biological function to randomly cloned DNA fragments.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00424421
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  • 10
    Electronic Resource
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    A yeast strain producing lys2 deletions at a high frequency after sporulation (1988)
    Springer
    Current genetics 14 (1988), S. 331-335 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Meiosis ; Deletion mutations ; Sequence dissimilarities
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A diploid yeast strain with extensive sequence dissimilarity in homologous regions near the LYS2 locus was sporulated, and spontaneous lys2 and lys5 mutant spores, selected on α-amino adipate, were analyzed. As many as 50% of the mutant spores contained a deletion in LYS2. These deletions occurred at a frequency of 5.0 × 10−7. While deletions of various sizes and endpoints were obtained, all the deletions recovered in this study included the border between homologous and non-homologous sequences located 4 kb upstream of LYS2. Large lys2 deletions that extended into an adjacent CYH2 duplication occurred at a frequency of 2.0 × 10−7, more than 1,000 times the frequency of the CYH2-LYS2 deletions found in a related haploid strain. This high frequency of CYH2-LYS2 deletions was observed only after sporulation of the diploid strain, and was dependent upon extensive sequence dissimilarity near the LYS2 locus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419990
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  • 11
    Electronic Resource
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    The allosuppressor gene SAL4 encodes a protein important for maintaining translational fidelity in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 537-543 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Allosuppressor ; Translation ; Fidelity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Allosuppressor (sal) mutations enhance the efficiency of the yeast ochre suppressor SUQ5 and define five unlinked loci, SALT-SALS. A number of sal4 mutants were isolated and found to have pleiotropic, allele;specific phenotypes, including hypersensitivity in vivo to paromomycin and other antibiotics that stimulate translational errors in yeast. To examine further the nature of the SAL4 gene product, the wild type SAL4 gene was isolated by complementation of a conditional lethal allele sal4-2, and demonstrated to be a single copy gene encoding a single 1.6 kb transcript. Restriction mapping and DNA hybridisation analysis were used to demonstrate that the SAL4 gene is identical to the previously identified omnipotent suppressor gene SUP45 (SUPT). Our results implicate the SAL4 gene product as playing a major role in maintaining translational accuracy in yeast.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00434078
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  • 12
    Electronic Resource
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    Hyperresistance to formaldehyde of Saccharomyces cerevisiae seems not to be correlated with the formation and removal of DNA protein cross-links (1988)
    Springer
    Current genetics 13 (1988), S. 125-128 
    Details
    ISSN: 1432-0983
    Keywords: Formaldehyde ; DNA-protein cross-links ; Repair ; Saccharomyces cerevisiae ; Hyperresistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The formation and removal of formaldehyde-mediated DNA protein cross-linking was measured by CsCI density gradient analysis in yeast strains of differing resistance to formaldehyde. Wild-type cells and transformants made hyperresistant to formaldehyde by a multi-copy vector containing the yeast SFA gene were specifically labeled in their DNA and incubated in the presence of formaldehyde. Treatment with formaldehyde lead to the formation of equal amounts of DNA protein cross-links; subsequent liquid holding of cells for 24 h resulted in the removal of nearly all DNA protein crosslinks regardless of the original formaldehyde resistance status of the strains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365646
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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
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    Yeast PAPS reductase: properties and requirements of the purified enzyme (1988)
    Springer
    Archives of microbiology 150 (1988), S. 313-319 
    Details
    ISSN: 1432-072X
    Keywords: 3′-Phosphoadenylyl sulphate reductase ; Sulphite formation ; Cysteine biosynthesis ; Thioredoxin ; Saccharomyces cerevisiae ; HPLC enzyme analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The enzymatic mechanism of sulphite formation in Saccharomyces cerevisiae was investigated using a purified 3′-phosphoadenylsulphate (PAPS) reductase and thioredoxin. The functionally active protein (MR 80–85 k) is represented by a dimer which reduces 3′-phosphoadenylyl sulphate to adenosine-3′,5′-bisphosphate and free sulphite at a stoichiometry of 1:1. Reduced thioredoxin is required as cosubstrate. Examination of the reaction products showed that free anionic sulphite is formed with no evidence for “bound-sulphite(s)” as intermediate. V max of the enriched enzyme was 4–7 nmol sulphite · min-1 · mg-1 using the homologous thioredoxin from yeast. The velocity of reaction decreased to 0.4 nmol sulphite · min-1 · mg-1 when heterologous thioredoxin (from Escherichia coli) was used instead. The K m of homologous thioredoxin was 0.6 · 10-6 M, for the heterologous cosubstrate it increased to 1.4 · 10-6 M. The affinity for PAPS remained practically unaffected (K m PAPS: 19 · 10-6 M in the homologous, and 21 · 10-6 M in the heterologous system). From the kinetic data it is concluded that the enzyme followed an ordered mechanism with thioredoxin as first substrate followed by PAPS as the second. Parallel lines in the reciprocal and a common intersect in the Hanes-plots for thioredoxin were seen as indication of a ping-pong (with respect to thioredoxin) uni-bi (with respect to PAPS) mechanism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00408300
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  • 15
    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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  • 16
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    Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation (1988)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
    Details
    ISSN: 1476-5535
    Keywords: Osmotic pressure ; Intracellular ethanol ; Yeast ; Nutrient ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The intracellular accumulation of ethanol in yeast and its potential effects on growth and fermentation have been topics of controversy for the past several years. The determination of intracellular ethanol based on the exclusion of [14C]sorbitol to estimate aqueous cell volume was used to examine the question of intracellular ethanol accumulation. An intracellular accumulation of ethanol inSaccharomyces cerevisiae was observed during the early stages of fermentation. However, as fermentation continued, the intracellular and extracellular concentrations of ethanol became similar. Increasing the osmotic pressure of the medium with glucose or sorbitol was observed to cause an increase in the intracellular ethanol concentration. Associated with this was a decrease in yeast growth and fermentation rates. In addition, increasing the osmotic pressure of the medium was observed to cause an increase in glycerol production. Supplementation of the media with excess peptone, yeast extract, magnesium sulfate and potassium phosphate was found to relieve the detrimental effects of high osmotic pressure. Under these conditions, though, no effect on the intracellular and extracellular ethanol distribution was observed. These results indicate that nutrient limitation, and not necessarily intracellular ethanol accumulation, plays a key role during yeast fermentations in media of high osmolarity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569575
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  • 17
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    Gating behaviors of a voltage-dependent and Ca2+-activated cation channel of yeast vacuolar membrane incorporated into planar lipid bilayer (1988)
    Springer
    The journal of membrane biology 106 (1988), S. 47-55 
    Details
    ISSN: 1432-1424
    Keywords: vacuole ; lipid bilayer ; K-channel ; single channel ; DIDS ; yeast ; Saccharomyces cerevisiae ; Ca2+ activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary A voltage-dependent and Ca2+-activated cation channel found in the vacuolar membrane of the yeast,Saccharomyces cerevisiae, was incorporated into planar lipid bilayer and its gating characteristics were studied at the macroscopic and single-channel levels. The open-channel probability at steady state, which was estimated by the macroscopic current measurement, gave a maximum value at −10 mV and decreased in a graded fashion as the voltage became more positive or more negative. The steady-state voltage dependence was explained by assuming two independent gates, which had different rate constants and opposite voltage dependence. The fast-responding gate opened when the voltage of thecis side (the side to which the vesicles were added) was made more negative and the slow-responding gate behaved in the opposite direction. Relatively high concentrations of Ca2+, about 1mm, were required on thecis side for opening the slow gate in a voltage-dependent manner. DIDS increased the open-channel probability of the fast gate when added to thecis side, but was ineffective on the slow gate.
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    URL: http://dx.doi.org/10.1007/BF01871766
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  • 18
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    Enhancement of copper resistance and CupI amplification in carcinogen-treated yeast cells (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 88-94 
    Details
    ISSN: 1617-4623
    Keywords: CupI ; Gene amplification ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Carcinogen-induced amplification at the CupI locus, coding for a metallothionein protein, was studied in the yeast Saccharomyces cerevisiae. Exposure of cells from three different haploid strains, 4939, DBY746 and 320, to chemical carcinogens such as N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), ethylmethanesulfonate (EMS) and 4-nitroquinoline-N-oxide (4NQO) enhanced the frequency of copper-resistant colonies up to several hundred fold. Copper-resistant clones obtained from strains DBY746 and 320, which contain more than one copy of the CupI locus, displayed a four-to eightfold amplification of the CupI sequences. In these clones the amplified CupI sequences were organized in a tandem array. Carcinogen treatment of strain 4939 in which only one copy of the CupI gene is present produced resistant colonies without CupI amplification. The possible use of the yeast system to study gene duplication and amplification is discussed.
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    URL: http://dx.doi.org/10.1007/BF00338397
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  • 19
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    Structural analysis of the 5′ regions of yeast SUC genes revealed analogous palindromes in SUC, MAL and GAL (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 446-454 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Invertase genes ; Promoter sequences ; Palindromes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the yeast Saccharomyces cerevisiae six unlinked structural genes for invertase, the SUC genes, are known. We sequenced about 800 bp of the 5′ non-coding region and the first 220 bp of the coding region of the genes SUC1, SUC3, SUC4 and SUC5 and compared them with the previously sequenced genes SUC2 and SUC7 (Sarokin and Carlson 1985a). All are highly homologous within the coding region but in the non-coding region SUC1 shows some differences and SUC2 is more highly diverged. Two different kinds of TATA boxes were identified: the more strongly expressed genes SUC1, 2 and 4 have the sequence TATAAA and the more weakly expressed genes SUC3, 5 and 7 have TACAAA. Though the SUC1 sequence is in general more homologous to the other SUC genes, the region between-140 and + 100 of SUC1 is nearly identical to SUC2. This could be due to a gene conversion between SUC1 and the silent suc2 o allele which occurs in the strains carrying SUC1. Within the upstream regions of all the SUC genes three regions with palindromic sequences analogous to stem and loop structures were identified. Comparable structure could be detected in similar positions in the upstream sequences of the divergently transcribed yeast gene pairs MAL6S-MAL6T and GAL1-GAL10. Implications for the importance of these structures in the regulation and initiation of transcription are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425699
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  • 20
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    The Escherichia coli proB gene corrects the proline auxotrophy of Saccharomyces cerevisiae pro1 mutants (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 124-128 
    Details
    ISSN: 1617-4623
    Keywords: l-azetidine-2-carboxylate resistance ; Escherichia coli ; γ-glutamyl kinase ; Proline ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We constructed plasmids carrying the Escherichia coli proB gene that encodes γ-glutamyl kinase, under the control of the yeast GAL1 promoter. This construction was carried out with both the wild-type proB + gene and a mutant allele, proB74, that specifies an enzyme resistant to feedback inhibition by proline. Yeast pro1 mutants harboring these plasmids are proline prototrophs. We conclude that the pro1 mutation results in a deficiency in the γ-glutamyl kinase activity in Saccharomyces cerevisiae. Expression of the proB74 allele in yeast resulted in enhanced resistance to the proline analogue l-azetidine-2-carboxylate and in a 2.4-fold elevation of the intracellular free proline levels. This result suggests that γ-glutamyl kinase is the rate limiting step in proline biosynthesis in yeast.
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    URL: http://dx.doi.org/10.1007/BF00322454
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  • 21
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    Random cloning of bent DNA segments from Saccharomyces cerevisiae and primary characterization of their structures (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 249-256 
    Details
    ISSN: 1617-4623
    Keywords: Bent DNA ; DNA structure ; Saccharomyces cerevisiae ; 2 μm circle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Until recently it was assumed that any short segment of DNA could be approximated as a straight rod. Many instances, however, have been reported in which the helical axis is curved. We have devised a simple method for selective identification of DNA segments containing a sequence-directed bend (curvature), by means of a two-dimensional polyacrylamide gel electrophoresis. In order to gain general insights into the structural features and the functional significance of sequence-directed bends, a bank of plasmids carrying bent DNA inserts from the Saccharomyces cerevisiae total genomic DNA was constructed. Primary characterizations of a set of bent DNA segments randomly cloned from S. cerevisiae are presented. One of the cloned DNA segments appears to be derived from a yeast plasmid, the 2 μm circle DNA.
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    Expression of MFα1 in MATa cells supersensitive to α-factor leads to self-arrest (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 85-88 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; α-Factor ; Cell-cycle arrest ; STE genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary MATa cells of Saccharomyces cerevisiae defective in both the SST1 and SST2 gene products exhibit selfarrest when they express the MFα1 gene under the control of the GAL1 promoter. This reponse to endogenously produced pheromone can be alleviated by mutations which prevent the production of, or response to, α-factor. Suppressors of the self-arrest phenotype include a class of mutants which remain responsive to low levels of pheromone, but are resistant to high levels of α-factor. One of these mutants has been mapped to chromosome X, 31 cM distal to SUP4, and defines a new locus designated STE18.
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    URL: http://dx.doi.org/10.1007/BF00340184
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    PHO85, a negative regulator of the PHO system, is a homolog of the protein kinase gene, CDC28, of Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 162-164 
    Details
    ISSN: 1617-4623
    Keywords: CDC28 ; Phosphate regulation ; PHO85 ; Protein kinase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The product of the PHO85 gene, which encodes one of the negative regulatory factors of the PHO system in Saccharomyces cerevisiae, shows significant amino acid sequence homology with the CDC28 protein kinase. However, overexpressing PHO85 did not suppress the temperature sensitive phenotype of the cdc28-1 mutation. The nucleotide sequence of the PHO85 gene strongly suggests the presence of an intron near the sequence encoding the N-terminal region.
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    URL: http://dx.doi.org/10.1007/BF00340196
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    Structure of the ARO3 gene of Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 165-169 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; ARO3 gene ; DAHP synthase ; DNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, the genes ARO3 and ARO4 encode isoenzymes of 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase. Both genes are derepressed seven-fold under the general control of amino acid biosynthesis. A previously isolated 1.7kb fragment containing the ARO3 gene and the 5′- and 3′-flanking regions was sequenced. The endpoints of the ARO3 transcript coding for a 370 amino acid protein were mapped by primer extension experiments and S1 nuclease digestion. Promoter elements involved in transcription initiation and responsible for the strong general control derepression response are discussed.
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    URL: http://dx.doi.org/10.1007/BF00340197
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    Structure of the Saccharomyces cerevisiae URA4 gene encoding dihydroorotase (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 134-141 
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    ISSN: 1617-4623
    Keywords: Dihydroorotase ; URA4 ; Saccharomyces cerevisiae ; DNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The URA4 gene of Saccharomyces cerevisiae, coding for the third enzyme of the pyrimidine pathway, has been cloned through phenotypic complementation of a ura4 mutant of S. cerevisiae. Subcloning of an original 9 kb DNA fragment, carrying the yeast URA4 gene, allowed us to localize the gene on a 2 kb ClaI-BamHI fragment. The sequence of the URA4 structural gene and surrounding DNA was determined by the dideoxynucleotide chain termination method. The URA4 gene encodes a dihydroorotase subunit of calculated molecular weight 40600. S1 nuclease mapping indicated that transcription of URA4 is initiated at four major start sites located at positions-42,-30,-22 and-18. A set of potentially significant sequences was identified in the 5′ OH non-coding region of the gene. The deduced amino acid sequence of dihydroorotase was examined and compared with homologous amino acid sequences of Salmonella typhimurium, Escherichia coli and Drosophila melanogaster. S. cerevisiae dihydroorotase shows 40% homology with the S. typhimurium and E. coli enzymes and 23% homology with the D. melanogaster enzyme. A potential active site has been predicted for dihydroorotase from these comparisons.
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    URL: http://dx.doi.org/10.1007/BF00322456
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    The sporulation capable (sca) mutation of Saccharomyces cerevisiae is an allele of the SIR2 gene (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 430-434 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; SCA gene ; RME1 gene ; Haploid meiosis ; Mating type
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have used the special properties of the spo13-1 mutation in order to study the regulation of yeast meiosis by the mating type loci. We have found that both the rme1-1 mutation and the sca mutation allow haploid meiosis in spo13-1 strains. Therefore, haploid meiosis is regulated in the same manner as diploid meiosis. Unlike rme1-1, the sca mutation allows meiosis through derepression of the silent mating type cassettes; sca strains can sporulate only because they express both MAT a and MATα information. We have found further that sca is an allele of SIR2, one of the genes involved in repression of the silent cassettes. Therefore, the RME1 gene is the only known candidate for a master negative regulator through which the MAT locus controls meiosis.
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    URL: http://dx.doi.org/10.1007/BF00425696
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    An AT rich region of dyad symmetry is a promoter element in the yeast TRP1 gene (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 472-476 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; TRP1 promoter ; REgion of dyad symmetry ; AT rich tracts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transcription from the yeast TRP1 promoter results in two classes of transcript, I and II, that are influenced by different promoter elements. The 5′ flanking region contains a region of dyad symmetry (RDS) which contains a 12 nucleotide AT rich inverted repeat, separated by a 21 bp spacer region. The RDS lies within a region of the promoter required for transcription of calls II RNAs. A series of internal deletions and insertions have been constructed in vitro around the RDS and the effect of each mutation on transcription has been analysed. Deleting either of the repeats abolishes class II transcription and disruption of both repeats influenced the levels of the larger class I transcripts. Deletion of the spacer had no effect but increasing the length to 33 bp reduced transcription. These results show that the RDS is an important component of the TRP1 promoter, that both repeats must be preserved and that there is some constraint on the spacing of the repeats for maximal function.
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    The promoter of the β-glucosidase gene from Kluyveromyces fragilis contains sequences that act as upstream repressing sequences in Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 150-154 
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    ISSN: 1617-4623
    Keywords: β-Glucosidase ; Kluyveromyces fragilis ; Saccharomyces cerevisiae ; Upstream repressing sequence ; Gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The relationship between the promoter length of the Kluyveromyces fragilis β-glucosidase gene and the level of its expression in Saccharomyces cerevisiae was studied by gene fusion between deleted promoter fragments of various lengths and the promoterless β-galactosidase gene of Escherichia coli. The removal of a region from position-425 to-232 led to a tenfold increase in the expression of the gene. The same results were obtained for the reconstructed β-glucosidase gene with the same promoter length. It is likely that the deletion of this part of the promoter removes negative regulatory elements which are functional in Saccharomyces cerevisiae. This increase in activity is the main event which may explain the high increase in gene expression (60-fold) previously observed for an upstream deletion obtained during subcloning experiments of the β-glucosidase gene. It is also shown that the expression of the gene greatly depends upon the nature of the recipient strain, the growth phase of the cell and that of the vector carrying it.
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    URL: http://dx.doi.org/10.1007/BF00333412
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    The detection of mitotic and meiotic aneuploidy in yeast using a gene dosage selection system (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 10-18 
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    ISSN: 1617-4623
    Keywords: Aneuploidy ; Yeast ; Saccharomyces cerevisiae ; Methyl benzimidazol-2-yl carbamate ; Mitosis ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A system is described in which spontaneous and chemically-induced mitotic and meiotic hyperploidy can be assayed in the same diploid culture of Saccharomyces cerevisiae. Monitoring gene dosage changes at two loci on chromosome VIII, the test utilizes a leaky temperature-sensitive allele arg4-8 and low level copper resistance conferred by the single copy allele cup1 s. An extra chromosome VIII provides simultaneous increased dosage for both genes, resulting in colonies that are both prototrophic for arginine at 30° C and copper resistant. During mitotic cell divisions in diploids, spontaneous chromosome VIII hyperploids (trisomes and tetrasomes) occur at a frequency of 6.4×10-6 per viable cell. Among ascospores, the spontaneous chromosome VIII disome frequency is 5.5×10-6 per viable spore. The tubulin-binding reagent methyl benzimidazol-2-yl carbamate (MBC) elicits enhanced levels of mitotic and meiotic aneuploidy relative to control levels. The system represents a novel model for examining chromosome behavior during mitosis and meiosis and provides a sensitive and quantifiable procedure for examining chemically induced aneuploidy.
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    Regulation of the TRP4 gene of Saccharomyces cerevisiae at the transcriptional level and functional analysis of its promoter (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 168-175 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; TRP4 gene ; PRtransferase ; Promoter analysis ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The TRP4 gene of Saccharomyces cerevisiae, which encoded anthranilate phosphoribosyl transferase (E.C.2.4.2.18), is subject to the general control of amino acid biosynthesis. The regulation takes place at the transcriptional level by increasing the amount of initiation and not by changing the stability of mRNA. We have observed a change in the utilization of TRP4 mRNA start sites, depending on whether cells were grown under repressing or derepressing conditions. The function of promoter elements has been tested by deletion analysis with a plasmid-encoded TRP4 gene. A routinely practicable method was used for copy-number calibration of plasmids based on 2 μm DNA. Promoter structures and spacing problems in the TRP4 promoter region are discussed.
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    Amplification of plasmid copy number by thymidine kinase expression in Saccharomyces cerevisiae (1988)
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    Molecular genetics and genomics 211 (1988), S. 155-159 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Yeast ; Copy number ; Thymidine kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2 μm circle-based chimaeric plasmid containing the yeast LEU2 and the Herpes Simplex Virus type 1 thymidine kinase (HSV-1 TK) genes was constructed. Transformants grown under selective conditions for the LEU2 gene harboured the plasmid at about 15 copies per cell whilst selection for the HSV-1 TK gene led to an increase to about 100 copies per cell. Furthermore, the plasmid copy number could be controlled by the stringency of selection for the TK gene, and the increase in TK gene dosage was reflected in an increase in intracellular thymidine kinase activity. The mitotic stability of the plasmid in “high-copy” and “low-copy” number cells was determined. “High-copy” number cells showed a greater mitotic stability. The relationship of TK expression to plasmid copy number may be useful for the isolation of plasmid copy number mutants in yeast and the control of heterologous gene expression.
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    The 2 μm D region plays a role in yeast plasmid maintenance (1988)
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    Molecular genetics and genomics 212 (1988), S. 426-431 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; 2 μm circle ; Plasmid-partitioning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast 2 μm circle encodes four major transcribed open reading frames, A, B, C and D. Products of ORF's A, B and C, together with the inverted repeats and the other cis-acting loci ORI and STB, have been shown to be involved in plasmid maintenance. However, the function of ORF D has remained unclear. We have therefore carried out studies on 2 μm derivatives with both insertional and frameshift mutations in D. Our results indicate that there is a protein product encoded by ORF D, which is involved in plasmid maintenance. When the copy number of the C gene was reduced to one, by chromosomal integration, we observed striking differences in the efficiency of partitioning of D + and D − plasmid derivatives. Absence of D function could be compensated by an increase in dosage of the C gene, indicating that the D product may act to regulate C expression. Since the C product has been implicated in copy number control as well as partitioning, our data suggest that the D product may also be involved in both of these processes.
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    A consensus transcription termination sequence in the promoter region is necessary for efficient gene expression of the TRP1 gene of Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 495-504 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; TRP1 gene ; Yeast promoter ; Yeast vectors ; Copy number
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The TRP1 gene of Saccharomyces cerevisiae is the only TRP gene which is not derepressible by the general control regulatory system. In the TRP1 promoter transcription starts at five initiation sites, organized in two clusters. The two transcripts of the first, more upstream cluster include a long leader sequence of approximately 200 bp. A transcriptional terminator element located in the 5′ region of the TRP1 gene is essential for accurate gene expression. In partial TRP1 promoters lacking the terminator, like the original EcoRI TRP1 fragment used in numberous vectors, plasmid-encoded transcription is initiated predominantly in adjacent vector regions, resulting mainly in large, poorly translated transcripts. This poor translation is not due to mRNA instability. The effect can be suppressed by introducing artificial transcription barriers between vector sequences and the truncated EcoRI TRP1 fragment.
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    Nuclear and mitochondrial revertants of a yeast mitochondrial tRNA mutant (1988)
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    Molecular genetics and genomics 213 (1988), S. 425-434 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Mitochondria ; Transfer RNA ; syn - mutation ; Revertants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We isolated revertants capable of respiration from the respiratory deficient yeast mutant, FF1210-6C/ 170, which displays greatly decreased mitochondrial protein synthesis due to a single base substitution at the penultimate base of the tRNAAsp gene on mitochondrial (mt) DNA. Three classical types of revertant were identified: (1) same-site revertants; (2) intragenic revertants which restore the base pairing in the acceptor stem of the mitochondrial tRNAAsp; and (3) extragenic suppressors located in nuclear DNA. In addition a fourth type of revertant was identified in which the mutant tRNAAsp is amplified due to the maintenance of both the original mutant mtDNA and a modified form of the mutant mtDNA in which only a small region around the tRNAAsp gene is retained and amplified. The latter form resembles the mtDNA in vegetative petite (rho -) strains which normally segregates rapidly from the wild-type mtDNA. Each revertant type was characterized genetically and by both DNA sequence analysis of the mitochondrial tRNAAsp gene and analysis of the quantity and size of RNA containing the tRNAAsp sequence. These results indicate that the mitochondrial tRNAAsp of the mutant retains a low level of activity and that the presence of the terminal base pair in tRNAAsp is a determinant of both tRNAAsp function and the maintenance of wild-type levels of tRNAAsp.
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    The base-alteration spectrum of spontaneous and ultraviolet radiation-induced forward mutations in the URA3 locus of Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 396-404 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; URA3 locus ; Cyclobutane dimer ; Pyrimidine-pyrimidone (6-4) photoproduct ; “A rule”
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A forward mutation system has been developed to obtain rapidly clonable mutants at the URA3 locus in yeast by means of selection for 5-fluoroorotic acid resistance. We have used this system to determine base changes in 35 spontaneous and 34 ultraviolet radiation-induced ura3 base substitution mutants. Other mutants (frameshift, deletion, duplication, replacement) were detected as well. Evidence is reported which suggests cyclobutane dimers are the principal mutagenic lesions induced by UV radiation in stationary phase cells of the yeast Saccharomyces cerevisiae. Since most of the induced lesions are at 5′-TT-3′ sites, the results suggest that the “A-rule”, preferential insertion of adenine residues opposite poorly pairing sites in DNA, does not apply for yeast cells irradiated in stationary phase, whereas the spontaneous mutation data indicate that the A-rule applies for cells in logarithmic phase. Most of the spontaneous mutations are transversions. UV-induced transitions and transversions occur at approximately equal frequencies.
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    Transmission of yeast mitochondrial loci to progeny is reduced when nearby intergenic regions containing ori sequences are deleted (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 425-432 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Mitochondrial DNA ; Intergenic sequences ; Transmission ; Genetic crosses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitochondrial DNAs (mtDNA) from four stable revertant strains generated from high frequency petite forming strains of Saccharomyces cerevisiae have been shown to contain deletions which have eliminated intergenic sequences encompassing ori1, ori2 and ori7. The deleted sequences are dispensable for expression of the respiratory phenotype and mutant strains exhibit the same relative amount of mtDNA per cell as the wild-type (wt) parental strain. These deletion mutants were also used to study the influence of particular intergenic sequences on the transmission of closely linked mitochondrial loci. When the mutant strains were crossed with the parental wt strains, there was a strong bias towards the transmission into the progeny of mitochondrial genomes lacking the intergenic deletions. The deficiency in the transmission of the mutant regions was not a simple function of deletion length and varied between different loci. In crosses between mutant strains which had non-overlapping deletions, wt mtDNA molecules were formed by recombination. The wt recombinants were present at high frequencies among the progeny of such crosses, but recombinants containing both deletions were not detected at all. The results indicate that mitochondrial genomes can be selectively transmitted to progeny and that two particular intergenic regions positively influence transmission. Within these regions other sequences in addition to ori/rep affect transmission.
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    Yeast nuclear gene CBS2, required for translational activation of cytochrome b, encodes a basic protein of 45 kDa (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 263-270 
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    ISSN: 1617-4623
    Keywords: DNA sequence ; PET gene ; Saccharomyces cerevisiae ; Transcription initiation ; Translation activator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In yeast, synthesis of apocytochrome b from mitochondrial COB mRNA depends on at least three nuclear gene products. The translation stimulatory effect by two of these nuclear genes, CBS1 and CBS2, is mediated by the 5′-untranslated leader of COB mRNA. In this report, we show that CBS2 is located on chromosome IV and provide genetic evidence that the CBS2 gene encodes a polypeptide. Determination of the DNA sequence reveals a contiguous open reading frame of 1167 bp. The deduced polypeptide has a calculated molecular weight of 44.5 kDa and is characterized by a high content of positively charged amino acids. It has no significant homology to any known protein. The CBS2 gene is transcribed into low abundance mRNA species with a major transcription initiation site located 97 bp upstream from the ATG start codon next to a poly(dA-dT) stretch.
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    The effect of growth conditions on production and excretion of extracellular antigens by three ascomycetous yeasts (1988)
    Springer
    Antonie van Leeuwenhoek 54 (1988), S. 235-244 
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    ISSN: 1572-9699
    Keywords: extracellular antigens ; extracellular polysaccharides ; Hansenula wickerhamii ; Saccharomyces cerevisiae ; Stephanoascus ciferrii ; yeast antigens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ascomycetous yeasts produce extracellular antigens that are almost specific for the species. The antigen production by Hansenula wickerhamii and Stephanoascus ciferrii was independent of the carbon source and was proportional to the final cell density of the cultures. The same was true of chemostat cultures of Stephanoascus ciferrii, irrespective of the dilution rate and whether glucose or ammonia was the limiting nutrient. In cultures of Saccharomyces cerevisiae, however, antigen excretion mainly took place in the late exponential growth phase. Large amounts of antigen were extracted from the cell wall of Saccharomyces cerevisiae. A small amount was detected in the cytoplasm.
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    α-factor enhancement of hybrid formation by protoplast fusion in Saccharomyces cerevisiae II (1986)
    Springer
    Current genetics 10 (1986), S. 943-945 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; α-factor ; Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When Mat a cells are treated with α-factor prior to being protoplasted and fused, the frequency of karyogamy is higher than in unarrested controls.
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    Genetic mapping of nuclear mucidin resistance mutations in Saccharomyces cerevisiae (1986)
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    Current genetics 10 (1986), S. 665-670 
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    ISSN: 1432-0983
    Keywords: Multiple drug resistance ; Genetic mapping ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the yeast Saccharomyces cerevisiae, two nuclear pleiotropic drug resistance mutations pdr3-1 (former designation muc PR) and pdr3-2 (former designation DRI9/T7) have been selected as resistant to mucidin and as resistant to chloramphenicol plus cycloheximide, respectively. The pdr3 mutations were found not to affect the plasma membrane ATPase activity measured in a crude membrane fraction. Meiotic mapping using strains with standard genetic markers revealed that mutation pdr3-1 is centromere linked on the left arm of chromosome II at a distance of 5.9 ± 3.3 cM from its centromere and 11.6 ± 3.1 cM from the marker pet9. The centromere linked pdr3-2 mutation exhibited also genetic linkage to pet9 with a map distance of 9.8 ± 3.2 cM. These results indicate that pdr3-1 and pdr3-2 are alleles of the same pleiotropic drug resistance locus PDR3 which is involved in the control of the plasma membrane permeability in yeast.
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    Identification and characterization of four new GCD genes in Saccharomyces cerevisiae (1986)
    Springer
    Current genetics 10 (1986), S. 657-664 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Amino acid biosynthesis ; General control ; GCD-genes ; GCN-genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutant strains, resistant against the amino acid analogues 5-methyltryptophan, 5-fluorotryptophan and canavanine were isolated, starting with a trp2 leaky auxotrophic strain. Of 10 such strains, only four turned out to be of the “general control derepressed” (gcd) mutant type. Three other isolates were shown to be defective in the general amino acid permease system, while the remaining three strains displayed low spore viability and were not further investigated. Complementation tests amongst the four new gcd-mutant strains, including strain RH558 gcd2-1 isolated earlier, yielded five complementation groups: GCD2, GCD3, GCD4, GCD5, and GCD6. All mutant strains showed a dual phenotype, which was not separable by wild type backcrosses: “constitutive derepression” and “slow growth”. Epistatis of all gcd mutations over gcn1-1, gcn2-1 and gcn3-1 was found with respect to both phenotypes, except for gcd5-1, which was lethal in these combinations. On the other hand gcn4-101 was found to be epistatic over all gcd mutations, but only with respect to the “constitutive derepression” phenotype, and not to “slow growth”; again the combination with gcd5-1 was lethal. Mutation gcd2-1 was mapped on chromosome VII, 50 cM from leu1 and 22 cM from ade6. A new model is discussed, in which GCD-genes are involved in the amino acid uptake into the vacuoles.
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    Genetic analysis of intergeneric hybrids obtained by protoplast fusion in yeasts (1986)
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    Current genetics 10 (1986), S. 491-494 
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    ISSN: 1432-0983
    Keywords: Protoplast fusion ; Saccharomyces cerevisiae ; Schwanniomyces castellii ; Starch fermentability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Prototrophic hybrids have been obtained by the fusion of various auxotrophic haploid strains of Saccharomyces cerepisiae and Schwanniomyces castellii. The fusion hybrids showed starch fermentability which derived from one of the fusion parents, S. castellii. Surprisingly, these fusion hybrids were found to exhibit excellent sporulation and spore germination. The progenies of these fusion hybrids showed a few aberrant segregations, but mostly normal segregation for auxotrophic genetic markers. They also showed many tetrads with an apparently digenic segregation (2:2, 3:1 and 4:0) for starch fermentation. On the other hand, mating types of segregants of the fusion hybrids were determined by the prototrophic recovery method. Consequently, tetrad types for mating type were mostly 2a:1α:1 non-mater and several asci showed tetrad types of 2a:2 non-mater and 2a:2α. The 60 prototrophic fusion hybrids and its segregants did not secrete α-amylase on the starch agar plate. However, all of the data suggested that fusion hybrid could carry two dominant genes (STAB and STAC) to ferment starch, and that the two genes STAB and STA2 may be identical or allelic as may be the genes STAC and STA3.
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    Chromosomal mapping of the uracil permease gene of Saccharomyces cerevisiae (1986)
    Springer
    Current genetics 11 (1986), S. 93-96 
    Details
    ISSN: 1432-0983
    Keywords: Uracil permease gene ; Saccharomyces cerevisiae ; Chromosomal mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The gene FUR4, coding for the uracil permease in Saccharomyces cerevisiae, was mapped on chromosome II, at a distance of 7.8 cM from the centromere on the right arm of the chromosome. In a first step, we used the chromosome loss mapping method developed by Falco and Botstein (1983) to determine on which chromosome the gene mapped. After the observation that FUR4 was closely linked to GAL10, one of the three genes forming the gal cluster (Bassel and Mortimer 1971), we could determine precisely the position of the gene on chromosome II.
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    Induction of yeast DNA ligase genes in exponential and stationary phase cultures in response to DNA damaging agents (1986)
    Springer
    Current genetics 11 (1986), S. 107-112 
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    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; DNA ligase ; DNA damage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary UV-irradiation of stationary phase cells of Saccharomyces cerevisiae and Schizosaccharomyces pombe leads to a 9-fold and 90-fold increase in transcript levels from the respective DNA ligase genes CDC9 and CDC17, whereas exponential cells show only 3-fold and 2-fold increases. Induction of CDC9 after MMS treatment and γ-irradiation was also observed by using a CDC9-lacZ translational fusion and assaying for β-galactosidase. Surprisingly, irradiation of S. cerevisiae induces only a 50% increase in DNA ligase itself, probably reflecting the extremely high in vivo stability of the enzyme. The UV-induction of ligase may be part of a “fail-safe” mechanism which, together with the enzyme stability, ensures adequate supplies of this essential enzyme.
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    Conserved and non-conserved features among the yeast T-y elements (1986)
    Springer
    Current genetics 11 (1986), S. 193-200 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Ty elements ; Transposable elements ; Retroviruses ; tRNA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated and characterized a Ty element from a yeast cosmid library which exhibits several unsual features: it is flanked by non-homologous delta elements and directly associated with a singular delta element. A tRNA(Glu3) gene and tRNA(Cys) gene are found in conjunction with this element, located in opposite orientation on either end of it. The sequence information now available for several Ty elements has been used in a detailed comparative analysis to determine conserved features among the Ty elements, preferably between class I elements and a class II element. Highly conserved sequence motifs appear to be located at the borders of particular segments that correspond to the putative protein domains of the Tys. Furthermore, we include a comparison of the best-conserved amino acid homologies for these putative proteins of Ty elements, transposable elements from other organisms and several retroviral proviruses to confirm their close structural resemblance.
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    URL: http://dx.doi.org/10.1007/BF00420606
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    Isolation and characterization of a conditional mutant in Saccharomyces cerevisiae producing rho° petites at the non-permissive temperature (1986)
    Springer
    Current genetics 11 (1986), S. 201-204 
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    ISSN: 1432-0983
    Keywords: Rho°-petites ; Lycorine ; Mitochondrial DNA replication ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper describes the isolation and characterization of mutants affected in the maintenance of the mitochondrial (mt) genome. The rationale of the screening procedure is based on the observation that the alkaloid lycorine inhibits growth of rho −-mutants, whereas rho°-mutants, devoid of mt DNA, are resistant to this drug (Del Giudice et al. 1984). Fourteen temperature sensitive mutants have been isolated that display the following phenotype: -Growth on fermentable medium at 23°C and 35°C (exclusion of general temperature-sensitive mutants). -no growth at 23°C and growth at 35°C on fermentable medium containing lycorine (selection for mutants producing rho°-petites). -growth at 23°C and no growth an 35°C on non-fermentable medium (selection for temperature-dependent loss of respiratory competence). These mutants were termed tmm (for temperature sensitive maintenance of mt genome). Mutant tmm1-1 was analyzed genetically and biochemically. It carries a recessive nuclear mutation which gives rise to 90–95% cytoplasmic petites at the non-permissive temperature. The population of petites consists of more than 95% rho°-petites as shown by their resistance to lycorine, by staining with 4′,6-diamidino-2-phenylindole (DAPI), and by Southern hybridization with mt DNA probes. Wild-type control cultures produced approximately 1% petites with less than 10% rho°-mutants.
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    Hyperresistance to DNA damaging agents in yeast (1986)
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    Current genetics 11 (1986), S. 211-215 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Hyperresistance ; DNA damaging agents ; Genotoxic effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In order to study resistance to DNA damaging agents, yeast DNA segments conferring hyperresistance in this organism to such genotoxic agents were selected for among yeast cells transformed by a yeast genome library based on the multi-copy vector plasmid YEp13. Genetic variants hyperresistant to 4-nitroquinohne-N-oxide, formaldehyde, and alkylating agents were isolated and the respective hyperresistance determinants shown to co-segregate with the vector plasmid. Phenotypical characterization indicated different degrees of resistance, few cases of cross-resistance and differing structural stability of the cloned DNA. By transfer to E. coli and subsequent retransformation of yeast a number of plasmids was shown to stably carry the genetic information for hyperresistance.
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    Cloning and expression on a multicopy vector of five invertase genes of Saccharomyces cerevisiae (1986)
    Springer
    Current genetics 11 (1986), S. 217-225 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Gene cloning ; Invertase genes ; Multicopy vector
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Six unlinked loci for invertase structural genes are known in the yeast Saccharomyces cerevisiae: SUC1-SUC5 and SUC7. These genes are similar in structure and expression but not identical. Different yeast strains possess none, one or several of these genes. We have isolated the genes SUC1-SUC5, subcloned them into the multicopy vector YEp24 and compared the expression of the five SUC genes in one recipient strain. SUC2 was isolated by transformation of a suc0 strain with a gene pool and complementation to sucrose fermentation. SUC4 was cloned from a minipool of chromosomal fragments which were shown to contain SUC4 by Southern hybridization. SUC1, SUC3 and SUC5 were isolated using the method of plasmid eviction. A plasmid containing regions flanking SUC4 was integrated next to these SUC genes. The plasmid together with the SUC genes were then cut out of the chromosome using an appropriate restriction endonuclease. The length of chromosomal DNA fragments containing the different SUC genes were 4.8 kb for SUC1, 5.2 kb for SUC2, 4.8 kb for SUC3, 12.8 kb for SUC4 and 17.2 kb for SUC5. Fragments containing the complete SUC genes and the sequences controlling their expression were subcloned into YEp24 and transformed into a strain without any active invertase gene. Invertase activity of transformants was measured after growth repressing (8% glucose) and derepressing (2% raffinose) conditions. As expected from results with strains carrying the individual SUC genes in a chromosomal location, the SUC genes were expressed to a different extent.
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    Isolation and genetical and biochemical characterization of mutants resistant to the alkaloid lycorine (1986)
    Springer
    Current genetics 11 (1986), S. 247-249 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Nucleo-mitochondrial interactions ; Mitochondrial status ; Dominant lycorine resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants resistant to 200 µg/ml of the alkaloid lycorine (LYC R) in non-fermentable substrate were isolated after nitrosoguanidine mutagenesis. Tetrad analysis and growth of heterozygous (LYC R/lyc s) diploids from two different mutants revealed that a single nuclear and dominant mutation is responsible for the resistant phenotype. In the wild type total protein synthesis is only slightly inhibited, whereas DNA and RNA synthesis is lowered to about 30% of the control. In the lycorine resistant mutants all macromolecular syntheses are unaffected by the drug.
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    The overproducing CYP1 and the underproducing hap1 mutations are alleles of the same gene which regulates in trans the expression of the structural genes encoding iso-cytochromes c (1986)
    Springer
    Current genetics 10 (1986), S. 339-342 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cytochrome c ; Regulatory gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The CYP1 gene has previously been identified as coding for a positive trans active factor that activates the expression of CYC1 and CYP3, which are the structural genes for isol- and iso2-cytochrome c. Two phenotypically distinct classes of CYP1 mutations can be obtained indicating that CYC1 and CYP3 are differentially regulated by the product of CYP1. The HAP1 gene codes for a product which has previously been proved to be necessary for the expression of the heme dependent CYC1-UAS1 cis regulatory sequence. In this article, we show by complementation and recombination that CYP1 and HAP1 are the same gene, moreover we identify hap1-1 as an iso2-cytochrome c underproducer mutation of the CYP1 gene.
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    Genetic characterization of an α-specific gene responsible for sexual agglutinability in Saccharomyces cerevisiae: mapping and gene dose effect (1986)
    Springer
    Current genetics 10 (1986), S. 353-357 
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    ISSN: 1432-0983
    Keywords: agα1 Mutant ; Agglutination ; Gene dose effect ; Mapping ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A recessive agα1 mutation leads to specific defect in sexual agglutinability specifically in α cells of the yeast Saccharomyces cerevisiae. The cryptopleurine resistance gene cryR 1, closely linked to the mating type locus, was used to select α/α strains which emerged from α/α strains by mitotic nonreciprocal recombination, to genetically analyse agα1, since agα1 is expressed only in α mating type. The agα1 gene was found to be linked to the centromere tightly, to met3 at 4.4 cM, and to ilv3 at 12 cM on chromosome X. Sexual agglutinability of α cells was shown to be dependent on the dose of the AGα1 gene, using α/α isogenic strains carrying AGα1/AGα1, AGα1/agα1 or agα1/agα1. The sst2-1 mutation did not suppress the agα1 mutation. Based on these results, function of the AGα1 gene is discussed.
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    Analysis of the energy metabolism after incubation of Saccharomyces cerevisiae with sulfite or nitrite (1986)
    Springer
    Archives of microbiology 145 (1986), S. 27-31 
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    ISSN: 1432-072X
    Keywords: Nitrite ; Sulfite ; Saccharomyces cerevisiae ; ATP ; Energy metabolism ; Inorganic phosphate ; Glyceraldehyde-3-phosphate dehydrogenase ; Glucose-6-phosphate dehydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract After addition of 5 mM sulfite or nitrite to glucose-metabolizing cells of Saccharomyces cerevisiae a rapid decrease of the ATP content and an inversely proportional increase in the level of inorganic phosphate was observed. The concentration of ADP shows only small and transient changes. Cells of the yeast mutant pet 936, lacking mitochondrial F1ATPase, after addition of 5 mM sulfite or nitrite exhibit changes in ATP, ADP and inorganic phosphate very similar to those observed in wild type cells. They key enzyme of glucose degradation, glyceraldehyde-3-phosphate dehydrogenase was previously shown to be the most sulfiteor nitrite-sensitive enzyme of the glycolytic pathway. This enzyme shows the same sensitivity to sulfite or nitrite in cells of the mutant pet 936 as in wild type cells. It is concluded that the effects of sulfite or nitrite on ATP, ADP and inorganic phosphate are the result of inhibition of glyceraldehyde-3-phosphate dehydrogenase and not of inhibition of phosphorylation processes in the mitochondria. Levels of GTP, UTP and CTP show parallel changes to ATP. This is explained by the presence of very active nucleoside monophosphate kinases which cause a rapid exchange between the nucleoside phosphates. The effects of the sudden inhibition of glucose degradation by sulfite or nitrite on levels of ATP, ADP and inorganic phosphate are discussed in terms of the theory of Lynen (1942) on compensating phosphorylation and dephosphorylation in steady state glucose metabolizing yeast.
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    Effect of anticalmodulin drugs on the action of yeast α factor pheromone (1986)
    Springer
    Archives of microbiology 145 (1986), S. 104-106 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; α Factor ; Trifluoperazine ; Chlorpromazine ; Calmodulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Saccharomyces cerevisiae α factor pheromone arrest growth of cells of the a mating type (MAT a) at the G1 phase of the cell cycle. When treatment of MAT a cells with α factor was carried out in the presence of anticalmodulin drugs, trifluoperazine or chlorpromazine, the extent of cell growth arrest induced by α factor was reduced or even became undetectable. These results lend support to the hypothesis that calmodulin plays a role as mediator in the action of α factor on MAT a cells.
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    Incorporation of mannoproteins into the walls of aculeacin A-treated yeast cells (1986)
    Springer
    Archives of microbiology 146 (1986), S. 214-220 
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    ISSN: 1432-072X
    Keywords: Yeasts ; Cell wall ; Mannoproteins ; Aculeacin A ; Glucan ; Protoplasts ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Inhibition of the synthesis of alkali-insoluble glucan by aculeacin A in Saccharomyces cerevisiae cells caused a decrease in the incorporation of a high molecular weight heterogeneous mannoprotein material and of a 33000 mannoprotein into the wall network. This was concomitant with the excretion of the latter molecule into the growth medium. Regenerating yeast protoplasts liberated considerable amounts of the heterogeneous material to the medium independently of the presence of aculeacin. The protoplast walls did lack this component and contained only minor amounts of the 33000 molecule, which was also completely absent from walls of aculeacin-treated protoplasts. Considerable levels of the 33000 species were immunodetected in the supernatants from treated and untreated protoplasts. These results point to the existence of specific interactions between the glucan network of the yeast cell surface and some of the wall mannoproteins. On the other hand, the presence of a population of SDS-solubilizable mannoproteins in the wall was independent of glucan levels.
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    Accumulation and secretion of exoglucanase activity in yeast secretory mutants (1986)
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    Archives of microbiology 146 (1986), S. 221-226 
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    ISSN: 1432-072X
    Keywords: Exoglucanase ; Saccharomyces cerevisiae ; Secretory mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Representative conditional yeast secretory mutants, blocked in transport of secretory and plasma membrane proteins from the endoplasmic reticulum (sec 18), from the Golgi body (sec 7) and in transport of secretory vesicles (sec 1), accumulated exoglucanase, a constitutive yeast activity, when incubated at the restrictive temperature (37°C). Different proportions of the accumulated activity were released by mutant cells under permissive conditions. The presence or absence of cycloheximide during the secretion period made no differences in the results. More than 90% of the internal activity was bound to membrane in wild type cells. However, only the soluble pool underwent changes during the accumulation or secretion periods. The bulk of secretory invertase accumulated by sec 1 was also soluble. By contrast sec 7 and sec 18 accumulated membrane-bound as well as soluble invertase forms and both were secreted in similar proportions in each mutant. More than 90% of the accumulated invertase was secreted at the permissive temperature in sec 18 cells. That percentage was significantly lower for exoglucanase (〈65%). Concomitantly, invertase accumulated by this mutant exited from the cells with a lower half time (t 1/2=150 min). These results may be interpreted assuming that exoglucanase is exported by a passive flow of the soluble pool.
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    Properties of the purified APS-kinase from Escherichia coli and Saccharomyces cerevisiae (1986)
    Springer
    Archives of microbiology 145 (1986), S. 32-38 
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    ISSN: 1432-072X
    Keywords: Sulphate assimilation ; Adenylylsulphate 3′-phosphotransferase EC 2.7.1.25 ; Escherichia coli ; Saccharomyces cerevisiae ; Enzyme purification ; Enzyme regulation ; Thiredoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Adenylylsulphate kinase (EC 2.7.1.25, ATP:adenylylsulphate 3′-phosphotransferase) has been isolated from Escherichia coli and from Saccharomyces cerevisiae. As major steps of purification, affinity chromatography on Sepharose CL 6B (“blue” or “red”) and chromatofocusing on polybuffer PBE 94tm were employed. The proteins were obtained in nearly homogeneous state after five chromatographic steps. The isolated enzymes from both sources appeared predominantly to exist as dimers. Upon reduction of the protein with dithiothreitol, it desintegrated into assumingly identical smaller subunits (E. coli rom Mr 90-85000 to 45-40000 and s. cerevisiae from 52-49500 to 28-29500). Both forms, dimer and monomer were found catalytically active. Preincubation of the isolated enzyme from either source in the presence of thioredoxin plus DTT, reduced glutathione or DTT increased the activity significantly. Treatment of the enzyme with SH-blocking reagents inactivated the enzyme irreversibly as compared to the inactivation caused by oxidants (2,6-dichlorophenol-indophenol, ferricyanide or oxydized glutathione). This oxidant induced inactivation was less pronounced for the fungal enzyme than for the bacterial protein. The enzyme from E. coli required thioredoxin in order to alleviate the GSSG-induced inactivation.
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    Turnover of abnormal proteins in Bacillus megaterium and Saccharomyces cerevisiae: differences between in vivo and in vitro degradation (1986)
    Springer
    Archives of microbiology 145 (1986), S. 97-103 
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    ISSN: 1432-072X
    Keywords: Bacillus megaterium ; Saccharomyces cerevisiae ; Ethionine ; Protein degradation ; Abnormal protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Degradation of abnormal proteins in Bacillus megaterium and Saccharomyces cerevisiae in vivo was compared with that in cell-free extracts. Protein degradation in vivo, when the cells were labelled with 14C-leucine during growth in the presence of ethionine, was affected by the concentration of the analogue used. Proteins synthesized in the presence of 0.2–1 mM ethionine were degraded most rapidly in both organisms. The proteolytic enzyme system of yeast degraded the analogue-containing proteins in vitro faster than the normal proteins. This holds also for proteins synthesized in the presence of 5 mM ethionine, whose degradation in vivo was impaired. The proteolytic system of B. megaterium, on the other hand, was unable in vitro to differentiate between normal and abnormal proteins. Denatured proteins underwent preferential degradation over normal and ethionine-containing proteins.
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    Yeasts in molecular biology. Spheroplast preparation withCanadida utilis, Schizosaccharomyces pombe andSaccharomyces cerevisiae (1986)
    Springer
    Bioscience reports 6 (1986), S. 597-602 
    Details
    ISSN: 1573-4935
    Keywords: Candida utilis ; cellulase ; DNAse ; β-glucuronidase ; Hansenula jadinii ; protoplast ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; spheroplast ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Efficient preparation of spheroplasts fromCandida utilis, Saccharomyces cerevisiae, andSchizosaccharomyces pombe, using a purified mixture of enzymes fromTrichoderma harzianum, is described. Limitations of other methods, and differences between yeasts are demonstrated.
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    Heme control region of the catalase T gene of the yeast Saccharomyces cerevisiae (1986)
    Springer
    Molecular genetics and genomics 203 (1986), S. 73-78 
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    ISSN: 1617-4623
    Keywords: Catalase T ; CTT1 ; Saccharomyces cerevisiae ; Yeast ; Heme control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The 5′-flanking region of the Saccharomyces cerevisiae catalase T gene (CTT1) and the part of the gene coding for the N-terminus of catalase T were sequenced. 5′-Ends of transcripts of the region were located by S1 nuclease mapping and primer extension. To analyse control elements in the upstream region, a CTT1-lacZ gene fusion was constructed. Deletion analysis was carried out within a part of the 5′-flanking region showing homology to the upstream region of the yeast CYC1 gene. Like the CTT1 gene, this gene is controlled by heme, oxygen and glucose. The results obtained show that the CTT1 gene is positively controlled by heme. Tentative evidence has been obtained for the involvement of upstream sequences homologous to USA1 and USA2 of the CYC1 gene in heme control. Further, a negative site has been located between the upstream activator sites and the transcription start. Within this negative region a ten base-pair sequence was detected that shows high homology to a sequence located within a negative control region of the CYC1 gene and some homology to the negative control elements of the S. cerevisiae CAR1 and CAR2 genes.
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    Transcription of genes encoding enzymes involved in DNA synthesis during the cell cycle of Saccharomyces cerevisiae (1986)
    Springer
    Molecular genetics and genomics 204 (1986), S. 363-366 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Gene expression ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have examined the pattern of transcription exhibited by four genes in the dTTP biosynthetic pathway of Saccharomyces cerevisiae. Consistent with the results reported previously by Storms et al. (1984), the TMP1 (or CDC21) gene encoding thymidylate synthase was found to be transcribed in a periodic manner during the cell cycle with maximal mRNA levels occurring just prior to the onset of DNA replication. Three other genes in this pathway DCD1, DUT1 and DFR1 encoding dCMP deaminase, dUTP pyrophosphatase and dihydrofolate reductase, respecitively, exhibited relatively constant levels of transcription throughout the cell cycle. These results, particularly for DFR1, are in marked contrast with those obtained in other eukaryotic systems which have suggested that, in general, genes encoding enzymes involved in DNA precursor synthesis are subject to cell cycle regulation. Thus, periodic transcription is not a property common to all genes involved in DNA replication in this eukaryote.
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    DNase I sensitivity of the chromatin of the yeast SUC2 gene for invertase (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 422-427 
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    ISSN: 1617-4623
    Keywords: SUC2 gene ; Active chromatin ; DNase I hypersensitive sites ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNase I sensitivity of chromatin of the yeast SUC2 gene, which encodes two forms of invertase, has been studied both in the genome and in a multicopy plasmid carrying the gene and its flanking sequences. Whereas little if any difference in the DNase I sensitivity of the flanking regions was found between the repressed and the derepressed states, derepression of the gene was accompanied by a large increase in the sensitivity of the transcribed region. A well-defined DNase I hypersensitive site was found centered at ∼ 120 bp downstream from the end of the coding region. This site seems to be flanked in the 3′ non-coding region by strictly positioned nucleosomes, and the structure of this region changes upon derepression. In the 5′ non-conding region two DNase I hypersensitive sites have been found flanking the TATA box and a set of three closely spaced hypersensitive sites occurs in an upstream regulatory sequence. The structure of these latter sites depends on the on-off state of transcription.
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    Cloning of the ARO3 gene of Saccharomyces cerevisiae and its regulation (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 353-357 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; ARO3 gene ; DAHP synthase ; Regulation ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Regulation of the two isozymes of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHP synthase; EC 4.1.2.15) encoded by the genes ARO3 and ARO4 of Saccharomyces cerevisiae was studied. Both genes were shown to respond equally well to the general control of amino acid biosynthesis. Strains with mutations in these two genes were obtained by selecting first for a single aro3 mutation and afterwards for a double aro3 aro4 mutation. Gene ARO3, coding for the phenylanine-dependent isozyme of DAHP synthase was cloned on the 2 μm multicopy vector pJDB207 by complementation of mutation aro3-1 in yeast. The ARO3 gene, carried originally on a 9.6 kb BamHI fragment (plasmid pME541A), was subcloned on a 1.9 kb HindIII-XbaI fragment (plasmid pME543). A transcript of about 1.5 kb was shown to proceed from the HindIII towards the XbaI site. Expression from the 9.6 kb as well as from the 1.9 kb fragment was normal on a multicopy vector, since in both cases DAHP synthase levels of about 50-fold the wild-type level were observed.
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    Positive and negative regulation ofCAR1 Expression inSaccharomyces cerevisiae (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 170-175 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Arginase ; Regulation of gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We localized the chromosomal targets of several of the regulatory controls of expression of theCAR1 gene. Fusion tolacZ of several fragments of the 5′ non-coding region showed that induction ofCAR1 by arginine is positively regulated by the products of theARGR genes. The target lies upstream of another site where repression by the CARGRI molecule occurs. The latter control is not specific to arginine catabolism since it also affectsCYC-1 and indeed does not appear to involve arginine. The primary target of the two other regulatory allelesCARGRII andCARGRIII is not situated in the 5′ non-coding region. Deletion analysis supports the fusion data and confirms the order of the regulatory regions: 5′—nitrogen catabolite repression—activation by arginine—CARGRI-mediated repression—CAR1.
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    Gene structure in the histidine operon of Escherichia coli (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 42-47 
    Details
    ISSN: 1617-4623
    Keywords: Minicells ; Histidinol phosphatase ; Recombinant plasmids ; Salmonella typhimurium ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The bifunctional enzyme imidazoleglycerolphosphate dehydratase and histidinolphosphate phosphatase is encoded by the hisB gene. The fourth gene of the histidine operon, hisB, was cloned and mapped on a 2,300 base pair DNA fragment. In the present study we report the complete nucleotide sequence of the hisB gene of Escherichia coli. The gene is 1,068 nucleotides long and codes for a protein of 355 amino acids with an apparent molecular weight of 39,998 daltons. The protein product(s) of the hisB region of both Salmonella typhimurium and E. coli were identified by subcloning and expression in an in vitro translation system. In both organisms the hisB gene directed the synthesis of a single protein with an apparent molecular weight of 40,500 daltons, consistent with the data derived from the nucleotide sequence analysis.
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    An altered ribosomal protein in an edeine-resistant mutant of Saccharomyces cerevisiae (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 120-124 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Ribosomal protein ; Edeine resistant mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The r-proteins of an edeine-resistant mutant of Saccharomyces cerevisiae were compared to those of the wild-type strain by using two different two-dimensional electrophoretic techniques: (1) the Kaltschmidt-Wittmann method and, (2) the Kaltschmidt-Wittmann system, in the first dimension and the Na Dodecyl-SO4 system in the second. With the first technique, the results indicate that the patterns of basic ribosomal proteins are similar in the two strains. However, the pattern of acidic ribosomal proteins of the mutant revealed an additional protein band with respect to the normal one. Using the other technique, the patterns of basic and acidic ribosomal proteins of the mutant demonstrated a similarity to the corresponding pattern of the wild-type strain. The data disclose that an acidic ribosomal protein of the mutant may have two forms with different electrophoretic mobilities and similar molecular weights.
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    In vivo homologous recombination intermediates of yeast mitochondrial DNA analyzed by electron microscopy (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 421-428 
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    ISSN: 1617-4623
    Keywords: Recombination intermediates ; Mitochondrial DNA ; Electron microscopy ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To study the structure of in vivo mitochondrial DNA recombination intermediates in Saccharomyces cerevisiae, we used a deletion mutant of the wild type mitochondrial genome. The mtDNA of this petite is composed of a direct tandem repetition of an ∼4,600 pb monomer repeat unit with a unique HhaI restriction enzyme site per repeat. The structure of native mtDNA isolated from log phase cells, and mtDNA crosslinked in vivo with trioxsalen plus UVA irradiation, was studied by electron microscopy. Both populations contained crossed strand “Holliday” type recombination intermediates. Digestion of both non-crosslinked and crosslinked and mtDNA with the enzyme HhaI released X and H shaped structures composed of two monomers. Electron microscopic analysis revealed that these structures had pairs of equal length arms as required for homologous recombination intermediates and that junctions could occur at points along the entire monomer length. The percentage of recombining monomers in both non-crosslinked and trioxsalen crosslinked mtDNA was calculated by quantitative analysis of all the structures present in an HhaI digest. The relationship between these values and the apparent dispersive replication of mtDNA in density-shift experiments and mtDNA fragility during isolation is discussed.
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    Partial complementation of the UV sensitivity of E. coli and yeast excision repair mutants by the cloned denV gene of bacteriophage T4 (1986)
    Springer
    Molecular genetics and genomics 203 (1986), S. 163-171 
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    ISSN: 1617-4623
    Keywords: Excision repair ; denV gene ; UV radiation ; Saccharomyces cerevisiae ; Phage T4
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The denV gene of bacteriophage T4 was reconstituted from two overlapping DNA fragments cloned in M13 vectors. The coding region of the intact gene was tailored into a series of plasmid vectors containing different promoters suitable for expression of the gene in E. coli and in yeast. Induction of the TAC promoter with IPTG resulted in overexpression of the gene, which was lethal to E. coli. Expression of the TACdenV gene in the absence of IPTG, or the use of the yeast GAL1 or ADH promoters resulted in partial complementation of the UV sensitivity of uvrA, uvrB, uvrC and recA mutants of E. coli and rad1, rad2, rad3, rad4 and rad10 mutants of S. cerevisiae. The extent of denV-mediated reactivation of excision-defective mutants was approximately equal to that of photoreactivation of such strains. Excision proficient E. coli cells transformed with a plasmid containing the denV gene were slightly more resistant to ultraviolet (UV) radiation than control cells without the denV gene. On the other hand, excision proficient yeast cells were slightly more sensitive to killing by UV radiation following transformation with a plasmid containing the denV gene. This effect was more pronounced in yeast mutants of the RAD52 epistasis group.
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    Spontaneous amplification of yeast CEN ARS plasmids (1986)
    Springer
    Molecular genetics and genomics 204 (1986), S. 98-102 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Minichromosomes ; Centromere ; Copy number ; Mitotic stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transformation of Saccharomyces cerevisiae with several yeast CEN4 ARS1 plasmids containing the his3-Δ4 allele (as well as the URA3 and TRP1 markers) yielded His+ transformants at 0.1%–50% the frequency of Ura+ Trp+ transformants. Additional His+ derivatives arose on continuous growth of transformants originally scored as His- Ura+ Trp+. In all cases, the His+ phenotype was not due to plasmid or host mutations but invariably correlated with an up to 12-fold increase in plasmid copy number. On removal of selective pressure, the His+ phenotype was lost more readily than the Ura+ Trp+ markers, with a corresponding decrease in plasmid copy number. Also, the amplification did not decrease the mitotic loss rate of the Ura+ Trp+ markers. These results indicate that CEN ARS plasmids can be spontaneously amplified to higher levels than previously observed. However, when amplified, apparently not all copies exhibit the characteristic stability of CEN ARS plasmids.
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    Heterothallic mating type switching in Saccharomyces cerevisiae is RAD52 dependent (1986)
    Springer
    Molecular genetics and genomics 204 (1986), S. 496-504 
    Details
    ISSN: 1617-4623
    Keywords: Heterothallic mating type switching ; Saccharomyces cerevisiae ; RAD52 ; RAD3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mating type interconversion of the yeast Saccharomyces cerevisiae is mediated by intrachromosomal gene conversion. Whereas homothallic switching is initiated by an endonuclease that produces a DNA double-strand cut within MAT, heterothallic strains lack this activity. In order to identify functions essential for initiation and realisation of heterothallic switching, repair-deficient strains carrying the rad52 or the rad3 mutation were constructed and tested for spontaneous and induced heterothallic switching frequencies. The wild type RAD52 function is essential for spontaneous and induced switching as well as for the intrachromosomal crossing over which produces a deleted ring chromosome III. The rad3 mutation had almost no influence on spontaneous or X-ray induced switching, but it does reduce induction by ultraviolet radiation. The data are interpreted to indicate that heterothallic switching is accomplished via recombinogenic repair, perhaps of a double-strand break. The conversion event as well as the crossing over event leading to a change in mating type are equally affected by the rad52 mutation and therefore perhaps associated.
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    Deletion of the phosphoglucose isomerase structural gene makes growth and sporulation glucose dependent in Saccharomyces cerevisiae (1986)
    Springer
    Molecular genetics and genomics 204 (1986), S. 310-316 
    Details
    ISSN: 1617-4623
    Keywords: Gene replacement ; PGI1 deletion ; Glucose-6-P ; Glucose dependence ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The structural gene PG11 coding for phosphoglucose isomerase was replaced by the LEU2 gene in the genome of Saccharomyces cerevisiae. Plasmids carrying the LEU2 gene between genomic regions flanking the PG11 gene were constructed and used to transform a PGI1/pgi1 diploid strain. Stable transformants lacking the PGI1 allele were isolated. Southern analysis of their meiotic products showed that haploid strains with a deletion of 1.6 kb within the 2.2 kb PG11 coding region were viable. Thus, the PGI1 gene is not essential in yeasts. However, unlike pgi1 mutants with residual phosphoglucose isomerase activity, no growth was detected in the pgi1Δ haploid strains when fructose was supplied as sole carbon source. The wild-type growth rate could be restored by adding 0.1% glucose to the medium. Furthermore, pgi1 mutants with residual enzymatic activity grew very slowly on fructose-supplemented media containing up to 2% glucose. Strains carrying the deletion allele, however, failed to grow at glucose concentrations higher than 0.5%. Also the pgi1Δ strains did not grow in glucose as sole carbon source. On the other hand pgi1Δ/pgi1Δ diploid strains did not sporulate on the usual acetate medium. This defect could be alleviated by the addition of 0.05% glucose to the sporulation medium. Under these conditions the pgi1Δ mutants sporulated with an efficiency of 25% compared with the wild type. These results suggest that (a) the phosphoglucose isomerase reaction is the only step catalysing the interconversion of glucose-6-P and fructose-6-P, (b) glucose-6-P is essential in yeasts, and (c) the oxidation of glucose-6-P through the glucose-6-P dehydrogenase reaction is not sufficient to support growth in yeasts.
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    Mutations in the right boundary of Saccharomyces cerevisiae centromere 6 lead to nonfunctional or partially functional centromeres (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 305-311 
    Details
    ISSN: 1617-4623
    Keywords: Centromere ; In vitro mutagenesis ; Mitosis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Centromeres most likely consist of DNA (CEN DNA) interacting with specific proteins. In Saccharomyces cerevisiae a clear picture has emerged of a 120 bp sequence that is characteristic of CEN DNA. We have investigated the 25 bp centromere DNA element (CDEIII) that represents the right part of a CEN DNA. We showed using a series of mutants generated in vitro that the right most triple A of the consensus sequence TGT.T.TG.. TTCCGAA.....AAA participates in the assembly of a functional centromere and that no further sequences to the right are needed. Distance changes between the centre dyad TTCCGAA and the triple A have two effects: Addition of one base pair leads to a reduction, and addition of two or four base pairs to a loss of centromere function implying a participation of the centre dyad and the triple A region in protein binding. Indeed, a synthetic ologonucleotide of 39 bp containing CDEIII shows specific protein binding.
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    Biosynthesis of superoxide dismutase and catalase inSaccharomyces cerevisiae: effects of oxygen and cytochromec deficiency (1986)
    Springer
    Journal of industrial microbiology and biotechnology 1 (1986), S. 187-193 
    Details
    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Cytochromec ; Superoxide dismutase ; Catalase ; Oxyradicals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Two strains ofSaccharomyces cerevisiae were used to study the synthesis of superoxide dismutase. One strain (cytochromec-deficient) contained 5–10% of the normal amounts of total cytochromec, while the other strain was a wild type. The cytochromec-deficient mutant had lower specific growth rate, growth yield, and oxygen uptake than the wild type. The superoxide dismutase and catalase activities, in both strains, were significantly lower under anaerobic than under aerobic conditions. Furthermore, under aerobic conditions the mutant contained higher levels of superoxide dismutase than the wild type which may be attributed to the higher intracellular flux of superoxide radicals caused by the cytochromec deficiency. The mutant also showed a lower level of catalase which was due to glucose repression.
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    Der Einfluß von Cadmium, Zink, Blei und Quecksilber auf die Enzymaktivität beiSaccharomyces cerevisiae in vitro (1983)
    Springer
    European journal of nutrition 22 (1983), S. 205-212 
    Details
    ISSN: 1436-6215
    Keywords: Schwermetallwirkung ; Malatdehydrogenase ; Glutamatdehydrogenase ; Glycerinaldehyd-3-phosphatdehydrogenase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The difference between cadmium, zinc, lead, and mercury in regard of their effects on the activity of the enzymes tested is very slight. Concentrations higher than 10−5 M reduce significantly the activity of the enzymes, and concentrations of approximately 10−3 M inhibit it completely. An increase of the activity cannot be detected. The addition of combinations of cadmium, zinc, and lead results in a summing up of the toxic effects, whereas the interaction between mercury and the other three heavy metals shows a cumulative effect, which is appointed nearly completely by the heavy metal more toxic. The findings suggest that under in-vitro conditions there exists a direct interaction between the heavy metals and the enzymes.
    Notes: Zusammenfassung Die vier Schwermetalle Cadmium, Zink, Blei und Quecksilber unterscheiden sich in ihrer Wirkung auf die Aktivität der untersuchten Enzyme nur sehr wenig. Konzentrationen über 10−5 M vermindern die Enzymaktivität signifikant, und Konzentrationen von etwa 10−3 M unterbinden sie völlig. Eine Steigerung der Enzymaktivität läßt sich nicht feststellen. Die Zugabe von Cadmium-, Zink- und Bleikombinationen führt zu einer Addition der toxischen Effekte, während bei der Interaktion zwischen Quecksilber und den anderen drei Schwermetallen die Gesamtwirkung fast ausschließlich durch das stärker hemmende Schwermetall allein bestimmt wird. Die erhaltenen Ergebnisse lassen vermuten, daß es unter Invitro-Bedingungen zu einer direkten Wechselwirkung zwischen den Schwermetallen und den Enzymen kommt.
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    Nuclear association in yeast of a hybrid vector containing mitochondrial DNA (1983)
    Springer
    Current genetics 7 (1983), S. 165-166 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cephalosporium acremonium ; Mitochondrial hybrid vector ; Nuclear association
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hybrid vector pCP2, consisting of the bacterial plasmid pBR325, the nuclear gene Leu-2 of Saccharomyces cerevisiae and a fragment of mitochondrial DNA from Cephalosporium acremonium, was found to associate with the nucleus in a transformed strain of Saccharomyces cerevisiae. This was inducted by (1) efficient expression of the Leu-2 gene as evidenced by a short generation time on selective medium; (2) independence of Leu-2 gene expression from mitochondrial protein synthesis, since pCP2 was shown to replicate and to be expressed in petite mutants; (3) association of pCP2 with isolated DNA from nuclei as proved by transformation experiments with E. coli.
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    Mating factor dependence of G1 cell cycle mutants of Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 309-312 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; G1 cdc mutants ; tα-factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants in four G1 cdc strains of Saccharomyces cerevisiae were isolated which failed to show division arrest in the presence of α-factor. The cell cycle properties, terminal arrest morphology and mating competence of these mutants at the restrictive temperature were examined. The G1 specific arrest of the cdc 36 and cdc39 mutants is dependent upon the availability of an intact mating factor response system in Mat a cells. Cdc28 and cdc37 mutants exert their cell cycle blocks independently of the mating factor pathway. It is likely that the nature of the primary growth defect in cdc36 and cdc39 mutants is such that the α-factor pathway is activated in the absence of the pheromone at the restrictive temperature and that G1 arrest is a secondary consequence of a non-cycle specific event in such mutants.
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    Mitotic segregation of 2 μm-pbr322 chimaeric plasmids in yeast (1983)
    Springer
    Current genetics 7 (1983), S. 235-237 
    Details
    ISSN: 1432-0983
    Keywords: DNA replication ; Shuttle vectors ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitotic segregation of three 2 μm-pBR322 chimaeric plasmids (YEp6, YEp21, and YEp24) was studied in yeast. Each displayed a characteristic rate of loss: YEp6 was lost at approximately twice the rate of YEp21 and YEp24. The loss rates were not significantly increased when two chimaeric plasmids were coresident, nor was the endogenous 2 μm plasmid itself displaced. Therefore these plasmids appear to be compatible in yeast.
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    Analysis of the effect of radiation repair mutations on the DEL1 mutator region of Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 57-61 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; DEL1 ; rad ; ste7
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In DEL1 strains of the yeast, Saccharomyces cerevisiae, the iso-1-cytochrome c (CYC1) region is flanked on either side by Tyl elements in direct orientation which promote cyc1 deletions of the bracketed DNA in the haploid cell. In this study, we asked which genes might control this event by testing the possibility that the DEL1 mutation mechanism requires an enzyme (or enzymes) that is also utilized in the repair of damaged DNA. To this end, we independently coupled eight repair mutations, rad3–2, rad4–4, rad6–1, rad6–3, rad9–1, rev3–1, rad50–1, and rad51-1, toDEL1 and asked whether DEL1 was still functional. We found that none of these rad mutations significantly affects the mutation frequency of 10−6-10−5 established in DEL1 strains for the CYC1 locus. Furthermore, we determined that ste7, a temperature-sensitive sterile allele known to alter gene regulation in Ty-mediated mutations, is not required for DEL1 function. Finally, DEL1 is not temperature-sensitive at 23° or 37 °C.
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    Leucine biosynthesis in yeast (1983)
    Springer
    Current genetics 7 (1983), S. 369-377 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Isoenzymes ; Induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Tetrad analysis indicates that α-isopropylmalate synthase activity of yeast is determined by two separate genes, designated LEU4 and LEU5. LEU4 is identified as a structural gene. LEU5 either encodes another α-isopropylmalate synthase activity by itself or provides some function needed for the expression of a second structural gene. The properties of mutants affecting the biosynthesis of leucine and its regulation suggest that the expression of LEU1 and LEU2 (structural genes encoding isopropylmalate isomerase and β-isopropylmalate dehydrogenase, respectively) is controlled by a complex of a-isopropylmalate and a regulatory element (the LEU3 gene product). Similarities and differences between yeast and Neurospora crassa with respect to leucine biosynthesis are discussed.
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    Trehalose: Its role in germination of Saccharomyces cerevisiae (1983)
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    Current genetics 7 (1983), S. 393-397 
    Details
    ISSN: 1432-0983
    Keywords: Trehalose ; Glycogen ; Sporulation ; Germination ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants with specific lesions were used to differentiate between the functions of glycogen and trehalose in S. cerevisiae. Diploids which harbor the glc1/glc1 mutation depend upon the phosphorylated, less active form of glycogen synthase and show a more active, phosphorylated form, of the enzyme trehalase. These conditions are due to a lesion in the regulating subunit of the cAMP-dependent protein kinase. Such cells are unable to sporulate. Diploids which contain the sst1/sst1 mutation have normal glycogen metabolism but their trehalose-6-phosphate synthase is not active. Such strains sporulate but germination is poor and only one-spore tetrads are formed. These results confirm that glycogen is needed to trigger sporulation while trehalose plays a role in the germination process. Different systems, I and II, of trehalose accumulation were proposed. System I would require the UDPG-linked trehalose synthase, whereas system II would constitute an alternative pathway, specifically induced or activated by the expression of a MAL gene. The presence of system II in its constitutive form in the constructed diploids would favour trehalose synthesis during growth on glucose, however, it did not overcome the glycogen deficiency during sporulation nor the lack of trehalose for germination. It seems that only system I, namely trehalose 6-P-synthase, plays a role in the germination process.
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    Protein secretion in yeast: Two chromosomal mutants that oversecrete killer toxin in Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 449-456 
    Details
    ISSN: 1432-0983
    Keywords: Oversecretion mutants ; Protease defect ; Wall glucan defect ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two chromosomal mutations in yeast that result in oversecretion of the K1 killer toxin protein were examined. A recessive mutation in gene ski5 appears to lead to toxin oversecretion through a defect in a cell surface, PMSF-inhibited protease. A wild type killer strain degraded toxin following synthesis, and degradation could be partially prevented by addition of PMSF to the growth medium. The ski5 mutation caused an approximate ten fold oversecretion of toxin, similar to that seen in a PMSF-treated wild type culture, and no increased oversecretion in the presence of PMSF. The ski5 mutation caused oversecretion of other low molecular weight secreted proteins and appeared to oversecrete the α-factor pheromone, as judged by activity tests. The ski5 mutation was complemented by mutations in ski genes 1–4, and the mutant was not supersensitive to mating pheromones or K2 killer toxin. We also examined killer strains with a mutation in the nuclear gene krel which results in a defective (1→6)-β-D-glucan cell wall receptor for killer toxin. Such strains oversecrete toxin into the growth medium, but also, unexpectedly, oversecrete most other secreted proteins. The defect in (1→6)-β-D-glucan in these mutants appears to perturb the partitioning of secreted proteins between the cell wall and the medium.
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    Analysis of yeast DNA by alkaline filter elution (1983)
    Springer
    Current genetics 7 (1983), S. 427-431 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; DNA ; Alkaline elution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The method of analysis of DNA in mammalian cells by alkaline elution from filters (Kohn et al. 1974) was adapted for studies on yeast DNA. By this technique spheroplasts obtained from yeast cells are lysed on filters and single-stranded DNA fragments selectively eluted by alkaline solutions. The procedure was applied to monitor the occurrence of replication intermediates and production of DNA single-strand breakage by MMS, and its repair in growth medium.
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    The study of a rDNA replicator in Saccharomyces (1983)
    Springer
    Current genetics 7 (1983), S. 433-438 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Yeast transformation ; Yeast autonomously replicating sequences ; Ribosomal RNA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have previously demonstrated that the loss of Rcp-CEN3, a centromeric plasmid containing yeast rDNA autonomously replicating sequences (ARS) is as high as around 50% per generation for most yeast strains. In this study we have attempted to elucidate mechanisms underlying the high mitotic instability of Rcp-CEN3. For this purpose a tandem duplication of a rDNA ARS was constructed in Rcp-CEN3. The new plasmid having two ARSs possesses a markedly higher mitotic stability as compared to a monoARS Rcp-CEN3. The mitotic stability of this centromere-containing plasmid which has two replicators corresponds to the calculated value for the mitotic stability of two monoARS plasmids Rcp-CEN3 in given cells. Genetic analysis has demonstrated that both plasmids having one or two ARSs are maintained in the single copy state. These results demonstrate that the mitotic instability of centromeric plasmid Rcp-CEN3 carrying a rDNA ARS is associated with the absence of stringent control of replication from the rDNA ARS. A possible mechanism of replication of the chromosomal rDNA repeats in yeast is discussed in the light of this data.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00377608
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  • 83
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    Mating-type-specific cell cycle arrest and shmoo formation by α pheromone of Saccharomyces cerevisiae in Hansenula wingei (1983)
    Springer
    Archives of microbiology 136 (1983), S. 79-80 
    Details
    ISSN: 1432-072X
    Keywords: α Pheromone ; Cell cycle ; G1 arrest ; Hansenula wingei ; Saccharomyces cerevisiae ; Shmoo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cell cycle of a (5) mating type cells of Hansenula wingei was arrested in the G1 phase by α pheromone of Saccharomyces cerevisiae but not by α(21) pheromone of H. wingei, although both the α pheromones are known to induce sexual agglutination ability of a mating type cells of H. wingei. Cells of α mating type of H. wingei became shmooed or arrested in the G1 phase in response to neither a pheromone of H. wingei nor α pheromone of S. cerevisiae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00415615
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  • 84
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    Electronic Resource
    Anaerobic growth of Saccharomyces cerevisiae in the absence of oleic acid and ergosterol? (1983)
    Springer
    Archives of microbiology 134 (1983), S. 64-67 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Anaerobic growth ; Hungate technique ; Tween 80 ; Ergosterol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Saccharomyces cerevisiae, Nontrachet strain 522 was successfully grown anaerobically on various glucose concentrations in Yeast Nitrogen Base (YNB) medium (pH 3.5) prepared under an atmosphere of carbon dioxide (CO2). This growth occurred in the absence of Tween 80 and ergosterol. The medium, prepared using the Hungate technique for cultivation of strictly anaerobic bacteria, contained the reducing agent cysteine·HCl·H2O (0.03%). Anaerobic growth was stimulated by the addition of Tween 80 and ergosterol to the anaerobic medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00429409
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  • 85
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    Inhibition of bud initiation by ethyl N-phenylcarbamate results in meiotic development in the yeast Saccharomyces cerevisiae (1983)
    Springer
    Archives of microbiology 134 (1983), S. 171-174 
    Details
    ISSN: 1432-072X
    Keywords: Acetate growth medium ; Anti-microtubule agent ; Bud initiation ; Ethyl N-phenylcarbamate ; Meiosis ; Mitotic cell cycle ; Saccharomyces cerevisiae ; Sporulation induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When diploid cells of Saccharomyces cerevisiae were incubated in acetate growth media containing 2.5 mM ethyl N-phenylcarbamate (EPC), bud initiation was inhibited preferentially, and eventually overgrown, unbudded cells accumulated. During subsequent incubation, meiosis and ascospore formation occurred at high frequencies. The behavior of EPC-treated cells was essentially the same as that of cells transferred to a starvation sporulation medium. EPC thus has a pronounced effect on the mitotic growth of yeast cells, which leads to meiotic development. Our observations indicate that EPC has a decisive function in the initiation of meiosis in rich growth media.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00407753
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  • 86
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    Electronic Resource
    Nucleotide pools of growing, synchronized and stressed cultures of Saccharomyces cerevisiae (1983)
    Springer
    Archives of microbiology 135 (1983), S. 63-67 
    Details
    ISSN: 1432-072X
    Keywords: Nucleotide pools ; Continuous cultivation ; Synchronized growth ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High pressure liquidd chromatography has been used to study the acid soluble nucleotide pool of Saccharomyces cerevisiae under different conditions of growth. ATP, ADP, AMP, NAD, GTP, UTP, UDP, CTP, CDP, and UDP-sugars plus UMP could be separated and were found in concentrations higher than 0.1 μmol per g yeast cell dry weight (=detection limit). During glucose-limited continuous culture the levels of individual nucleotides depended on the growth rate, which was most pronounced with pyrimidine (uridine, cytidine) nucleotides. The energy charge (E.C.) remained high (0.9) at all growth rates (0.07–0.3 h-1). During synchronized growth at a constant growth rate (0.11 h-1) almost all nucleotide levels and the E.C. remained at constant values with the only exception of UDP-sugars and UMP of which increased levels were found during the phase of budding. Under conditions of metabolic stress (addition of antimycin A, deoxyglucose plus iodoacetate) pronounced changes in the levels of purine (adenine and guanine) nucleotides and the E.C. were observed. All other nucleotides were less influenced by these conditions. Only under these conditions IMP accumulation was observed. The results strongly argue against the significance of purine nucleotide or E.C. measurements under viable conditions. In contrast, changes in the levels of pyrimidine nucleotides seem to be indicative of changes in the flux through the metabolic pathways where they act as coenzymes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419484
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  • 87
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    Electronic Resource
    Interaction of ethidium bromide with yeast cells investigated by electron probe X-ray microanalysis (1983)
    Springer
    The journal of membrane biology 73 (1983), S. 131-136 
    Details
    ISSN: 1432-1424
    Keywords: electron probe X-ray microanalysis ; Saccharomyces cerevisiae ; ethidium ; brontophenol blue ; cationic dye ; cytolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary K+ efflux provoked by ethidium proceeds partially as an all-or-none effect by which the diffusion barrier for K+ is disrupted and partially from still intact cells, presumably by exchange against ethidium. This is shown by the application of an electron probe microanalysis X-ray technique by which the K+ content of a number of individual cells is analyzed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01870436
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