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  • Saccharomyces cerevisiae  (233)
  • Springer  (233)
  • Periodicals Archive Online (PAO)
  • 1985-1989  (169)
  • 1980-1984  (64)
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  • 1
    Electronic Resource
    Electronic Resource
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01871766
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  • 2
    Electronic Resource
    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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  • 3
    Electronic Resource
    Electronic Resource
    Isolation and characterization of a mutant of Saccharomyces cerevisiae defective in phosphoenolpyruvate carboxykinase (1982)
    Springer
    Archives of microbiology 132 (1982), S. 141-143 
    Details
    ISSN: 1432-072X
    Keywords: Phosphoenolpyruvate carboxykinase ; Gluconeogenesis ; Saccharomyces cerevisiae ; Mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mutant of Saccharomyces cerevisiae lacking phosphoenolpyruvate carboxykinase (E.C. 4.1.1.32) was isolated. The mutant did not grow on gluconeogenic sources except glycerol. The mutation was recessive and apparently affected the structural gene of the enzyme. Intracellular levels of metabolites related to the metabolic situation of the enzyme were not significantly affected after transfer of the mutant from a medium with glycerol to a medium with ethanol as carbon source. In these conditions only AMP decreased 3 to 5 times. A search for mutants affected in the other gluconeogenic enzyme, fructose 1,6 bisphosphatase, remained unsuccessful.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00508719
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  • 4
    Electronic Resource
    Electronic Resource
    Temperature dependency of induction of sexual agglutinability by α pheromone in the yeast Saccharomyces cerevisiae (1982)
    Springer
    Archives of microbiology 132 (1982), S. 236-240 
    Details
    ISSN: 1432-072X
    Keywords: α Pheromone ; Cycloheximide ; Inducible a strain ; Phenylmethylsulfonyl fluoride ; Saccharomyces cerevisiae ; Sexual agglutinability ; Temperature-sensitive
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When α pheromone-pretreated cells of an inducible a strain of Saccharomyces cerevisiae carrying the inducible gene saa1 were incubated in a growth medium at 28°C, induction of sexual agglutinability began after a 10 min lag period. If the cells were incubated at 38°C during the lag period, no induction occurred even after incubation at 28°C. Contrary to this, if the cells were incubated at 28°C during the lag period, almost complete induction occurred, even after transfer to 38°C. Temperature shift experiments revealed that 5 min incubation at 28°C was necessary for the initiation of the temperature-sensitive period and further 5 min incubation for the completion of the period. The temperature-sensitive period was sensitive to phenylmethylsulfonyl fluoride.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00407957
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  • 5
    Electronic Resource
    Electronic Resource
    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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  • 6
    Electronic Resource
    Electronic Resource
    α-mating-type-specific suppression and a-mating-type-specific enhancement by tunicamycin of sexual agglutinability in the yeast Saccharomyces cervisiae (1984)
    Springer
    Archives of microbiology 138 (1984), S. 310-314 
    Details
    ISSN: 1432-072X
    Keywords: Glycoprotein ; Inducible strains ; Saccharomyces cerevisiae ; Sexual agglutinability ; Tunicamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effects of tunicamycin (TM) on the sexual agglutinability and zygote formation of Saccharomyces cerevisiae were studied using the two kinds of haploid strains, inducible and constitutive for sexual agglutinability. Induction of sexual agglutinability by opposite mating type sex pheromone of inducible strains was inhibited by TM in α mating type but not in a mating type. The recovery by temperature-shift-down from the temperature-suppressed sexual agglutinability of constitutive strains was enhanced by TM in a mating type but rather inhibited in α mating type. Pretreatment with TM of constitutive strains enhanced sexual agglutinability in a mating type but not in α mating type. The above-mentioned a-mating-type-specific agglutinability-enhancing actions of TM were discussed in relation to the action mechanism of α pheromone which induces or enhances the sexual agglutinability of a cells. Zygote formation was inhibited by TM in both constitutive and inducible strains at concentrations which showed only partially inhibitory effect on sexual agglutinability.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00410896
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  • 7
    Electronic Resource
    Electronic Resource
    Properties of the purified APS-kinase from Escherichia coli and Saccharomyces cerevisiae (1986)
    Springer
    Archives of microbiology 145 (1986), S. 32-38 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00413024
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  • 8
    Electronic Resource
    Electronic Resource
    Analysis of the energy metabolism after incubation of Saccharomyces cerevisiae with sulfite or nitrite (1986)
    Springer
    Archives of microbiology 145 (1986), S. 27-31 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00413023
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  • 9
    Electronic Resource
    Electronic Resource
    The effect of sulfite on the yeast Saccharomyces cerevisiae (1980)
    Springer
    Archives of microbiology 125 (1980), S. 89-95 
    Details
    ISSN: 1432-072X
    Keywords: Sulfur dioxide ; Sulfite ; Air polluting substances ; Saccharomyces cerevisiae ; Active agent of irreversible cell inactivation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract After a short period of tolerance, living cells of Saccharomyces cerevisiae were irreversibly damaged by low concentrations of sulfite. The length of the period of tolerance and the rate of the damaging effect depended on the concentration on sulfite, pH-value, temperature, the physiological state of the cells, and incubation time. Inhibitors of protein synthesis and mitochondrial ATP synthesis did not alter the deleterious effect of sulfite on living cells. Furthermore, cell damage leading to inhibition of colony formation occured under aerobic as well as under anaerobic conditions. Prior to cell inactivation sulfite induced the formation of respiratory deficient cells. The active agent was shown to be SO2.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00403203
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  • 10
    Electronic Resource
    Electronic Resource
    Effect of growth temperature upon heat sensitivity in Saccharomyces cerevisiae (1980)
    Springer
    Archives of microbiology 124 (1980), S. 285-287 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Heat killing ; Membrane damage ; Genetic damage ; Growth temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The resistance of exponentially growing yeast cells to killing by exposure to 52°C increased markedly as the growth temperature was increased. Identical killing curves were obtained for cells suspended in growth medium or in 0.9% saline. Cells resistant to killing at 52°C were quite sensitive to killing at slightly higher temperatures. These results suggest a primary role for membrane damage in the mechanism of heat killing.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00427739
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  • 11
    Electronic Resource
    Electronic Resource
    The use of phenylmethylsulfonyl fluoride in the study of catabolite inactivation and repression in intact cells of Saccharomyces cerevisiae (1980)
    Springer
    Archives of microbiology 124 (1980), S. 293-295 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Catabolite repression and inactivation ; Inhibition of protease B
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Catabolite inactivation of fructose-1,6-bisphosphatase, isocitrate lyase, phosphoenolpruvate carboxykinase and malate dehydrogenase in intact cells could be prevented by phenylmethylsulfonyl fluoride added 40 min prior to the addition of glucose. Protein synthesis, fermentative and respiratory activity and catabolite repression were not affected. Elimination of catabolite inactivation by the addition of PMSF revealed that catabolite repression started at different times for different enzyme.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00427741
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  • 12
    Electronic Resource
    Electronic Resource
    The structural organization of the Tibi grain as revealed by light, scanning and transmission microscopy (1980)
    Springer
    Archives of microbiology 128 (1980), S. 157-161 
    Details
    ISSN: 1432-072X
    Keywords: Lactobacillus brevis ; Streptococcus lactis ; Saccharomyces cerevisiae ; Concanavalin A ; Symbiosis ; Tibi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tibi grains consist of a unique and very stable symbiotic association of Lactobacillus brevis, Streptococcus lactis and Saccharomyces cerevisiae embedded in a dextran matrix. The structural organization of the grain was examined by light, scanning and transmission electron microscopy. The grain was constituted of an outer compact layer and an inner spongy structure. The outer layer was more densely populated by the microorganisms than the inner layer but dextran, stained on frozen thin sections with fluorescein-conjugated concanavalin A, was more abundant in the inner layer.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00406153
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  • 13
    Electronic Resource
    Electronic Resource
    Concentration of metabolites and the regulation of phosphofructokinase and fructose-1,6-bisphosphatase in Saccharomyces cerevisiae (1981)
    Springer
    Archives of microbiology 129 (1981), S. 216-220 
    Details
    ISSN: 1432-072X
    Keywords: Fructose-1,6-bisphosphatase ; Phosphofructokinase ; Antagonistic enzymes ; Glycolytic intermediates ; ATP ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The intracellular levels of adenosine triphosphate and several glycolytic intermediates were determined in Saccharomyces cerevisiae in relation to the presence of the metabolically antagonistic enzymes phosphofructokinase and fructose-1,6-bisphosphatase. Phosphofructokinase is synthesized constitutively in cells grown in the presence of glucose and fructose-1,6-bisphosphatase derepression occurs upon the exhaustion of glucose from the growth medium. Transcriptional regulation of fructose-1,6-bisphosphatase was suggested based on experiments with wild type cells using 8-hydroxyquinoline, a known inhibitor of nuclear transcription, and with the S. cerevisiae mutant strain A364A (ts-136) blocked in the transport of nuclear RNA at non-permissive temperature. The level of phosphofructokinase was reduced more than 25-fold under conditions of high citrate accumulation in an aconitaseless, glutamate requiring mutant strain, MO-1-9B. There was a rapid decrease in the levels of adenosine triphosphate and fructose-1,6-bisphosphate at the end of log-phase of culture growth when both fructose-1,6-bisphosphatase and phosphofructokinase were present in the cells simultaneously. The changes in the levels of key glycolytic intermediates, but not the changes in adenosine triphosphate, during the simultaneous presence of these two enzymes, can be explained without involving any futile cycling.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425254
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  • 14
    Electronic Resource
    Electronic Resource
    Homoserine and threonine pools of borrelidin resistant Saccharomyces cerevisiae mutants with an altered aspartokinase (1981)
    Springer
    Archives of microbiology 129 (1981), S. 368-370 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Borrelidin ; Aspartokinase ; Feedback regulation ; Threonine pool ; Homoserine pool ; S-Adenosylmethionine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Borrelidin is a specific inhibitor of the threonyl-tRNA-synthethase. A class of dominant borrelidin resistant mutants (BOR1) of Saccharomyces cerevisiae was biochemically characterized. The mutants possess an altered aspartokinase which is insensitive to threonine inhibition. The threonine and homoserine pools in these mutants are 22 times larger than in the wild type. By feeding aspartate under a variety of conditions the homoserine pool is increased even 57 times whilst the threonine pool is reduced.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00406464
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  • 15
    Electronic Resource
    Electronic Resource
    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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  • 16
    Electronic Resource
    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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  • 17
    Electronic Resource
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    Utilization of formate as an additional energy source by glucose-limited chemostat cultures ofCandida utilis CBS 621 andSaccharomyces cerevisiae CBS 8066 (1985)
    Springer
    Archives of microbiology 142 (1985), S. 302-306 
    Details
    ISSN: 1432-072X
    Keywords: Candida utilis ; Saccharomyces cerevisiae ; Glucose ; Formate ; Mixed-substrate utilization ; Growth yield ; Transhydrogenase ; Continuous culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Candida utilis CBS 621 andSaccharomyces cerevisiae CBS 8066 were grown in glucose-limited chemostat cultures with formate as an additional energy source. In both yeasts formate was oxidized via a cytoplasmic NAD+-linked formate dehydrogenase. Other formate-oxidizing enzymes could not be detected. WithCandida utilis the steady-state cell yield on glucose increased with increasing amounts of formate in the medium until growth became carbon-limited. The maximum growth yield on glucose in the presence of excess formate was dependent on the nitrogen source used for growth. With ammonium and nitrate the maximum yields were 0.69 and 0.56 g cells/g glucose, respectively. Calculations showed that this difference correlates with the NADPH requirement for biomass formation with these two nitrogen sources. This implies that the NADH produced from formate oxidation cannot replace the NADPH needed for biomass formation. It therefore is concluded that inCandida utilis transhydrogenase activity is absent. AlsoSaccharomyces cerevisiae was capable of oxidizing formate in glucose-limited chemostat cultures. However, in contrast toCandida utilis utilization of formate by this yeast did not enhance the cell yield on glucose.
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    URL: http://dx.doi.org/10.1007/BF00693408
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  • 18
    Electronic Resource
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    Effects of m-Cl-peroxy benzoic acid on glycolysis in Saccharomyces cerevisiae (1985)
    Springer
    Archives of microbiology 143 (1985), S. 220-224 
    Details
    ISSN: 1432-072X
    Keywords: Peroxy benzoic acid ; Saccharomyces cerevisiae ; ATP ; Glycolysis ; Glyceraldehyde-3-phosphate dehydrogenase ; Colony forming capacity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Concentrations of m-Cl-peroxy benzoic acid (CPBA) higher than 0.1 mM decrease the ATP-content of Saccharomyces cerevisiae in the presence of glucose in 1 min to less than 10% of the initial value. In the absence of glucose, 1.0 mM CPBA is necessary for a similar effect. After the rapid loss of ATP in the first min in the presence of glucose caused by 0.2 mM CPBA, the ATP-content recovers to nearly the initial value after 10 min. Aerobic glucose consumption and ethanol formation from glucose are both completely inhibited by 1.0 mM CPBA. Assays of the activities of nine different enzymes of the glycolytic pathway as well as analysis of steady state concentrations of metabolites suggest that glyceraldehyde-3-phosphate dehydrogenase is the most sensitive enzyme of glucose fermentation. Phosphofructokinase and alcohol dehydrogenase are slightly less sensitive. Incubation for 1 or 10 min with concentrations of 0.05 to 0.5 mM CPBA causes a) inhibition of glyceraldehyde-3-phosphate dehydrogenase, b) decrease of the ATP-content and c) a decrease of the colony forming capacity. From these findings it is concluded that the disturbance of the ATP-producing glycolytic metabolism by inactivation of glyceraldehyde-3-phosphate dehydrogenase may be an explanation for cell death caused by CPBA.
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    URL: http://dx.doi.org/10.1007/BF00411239
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  • 19
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    Glycerol production in relation to the ATP pool and heat production rate of the yeasts Debaryomyces hansenii and Saccharomyces cerevisiae during salt stress (1987)
    Springer
    Archives of microbiology 147 (1987), S. 358-363 
    Details
    ISSN: 1432-072X
    Keywords: Salt stress ; Debaryomyces hansenii ; Saccharomyces cerevisiae ; Glycerol ; ATP pool ; Microcalorimetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Changes in glycerol production and two parameters related to energy metabolism i. e. the heat production rate and the ATP pool, were assayed during growth of Saccharomyces cerevisiae and Debaryomyces hansenii in 4 mM and 1.35 M NaCl media. For both of the yeasts, the specific ATP pool changed during the growth cycle and reached maximum values around 10 nmol per mg dry weight in both types of media. The levels of glycerol were markedly enhanced by high salinity. In the presence of 1.35 M NaCl, D. hansenii retained most of its glycerol produced intracellularly, while S. cerevisiae extruded most of the glycerol to the environment. The intracellular glycerol level of S. cerevisiae equalled or exceeded that of D. hansenii, however, with values never lower than 3 μmol per mg dry weight at all phases of growth. When D. hansenii was grown at this high salinity the intracellular level of glycerol was found to correlate with the specific heat production rate. No such correlation was found for S. cerevisiae. We concluded that during salt stress, D. hansenii possesses the capacity to regulate the metabolism of glycerol to optimize growth, while S. cerevisiae may not be able to regulate when exposed to different demands on the glycerol metabolism.
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    URL: http://dx.doi.org/10.1007/BF00406133
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  • 20
    Electronic Resource
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    Cell wall mannoproteins during the population growth phases in Saccharomyces cerevisiae (1987)
    Springer
    Archives of microbiology 148 (1987), S. 88-94 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Mannoproteins ; Glucan ; Cell wall ; Population growth cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mannoproteins from cell walls of Saccharomyces cerevisiae synthesized at successive stages of the population growth cycle have been solubilized with Zymolyase and subsequently analyzed. The major change along the population cycle concerned a large size mannoprotein material; the size of the newly-synthesized molecules varied from 120,000–500,000 (mean of about 200,000) at early exponential phase to 250,000–350,000 (mean of about 300,000) at late exponential phase. These differences are due to modifications in the amount of N-glycosidically linked mannose residues, since the size of the peptide moiety was 90,000–100,000 at all growth stages and the level of O-glycosylation changed only slightly. After, incubation of the purified walls with concanavalin A-ferritin and subsequent analysis by electron microscopy, labelling was localized at the external and internal faces of the walls. The middle space of these was labelled after digestion of the glucan network with Zymolyase, which demonstrate the presence of mannoproteins in close contact with the structural glucan molecules throughout the wall.
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    URL: http://dx.doi.org/10.1007/BF00425354
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  • 21
    Electronic Resource
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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.
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    Cloning of the LEU2 gene of Saccharomyces cerevisiae by in vivo recombination (1989)
    Springer
    Archives of microbiology 152 (1989), S. 263-268 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Recombination ; Tryptophan cluster ; Yeast vectors ; Plasmid copy number
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe a convenient method for the in vivo construction of large plasmids that possess a multitude of restriction sites. A large (23 kbases) circular self-replicating plasmid carrying a partial LEU2-d gene was cotransformed with a circular non-replicating plasmid carrying the entire LEU2 gene. In vivo recombination results preferentially in a plasmid that carries both the LEU2-d and the entire LEU2 gene. In addition we also found one plasmid with a tandem LEU2 insertion and one plasmid where the LEU2-d gene was replaced by the entire LEU2 gene.
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    Sex pheromone of α mating type in the yeast Saccharomyces kluyveri and its synthetic analogues in relation to sex pheromones in Saccharomyces cerevisiae and Hansenula wingei (1982)
    Springer
    Archives of microbiology 132 (1982), S. 225-229 
    Details
    ISSN: 1432-072X
    Keywords: a Pheromone ; α Pheromone ; Hansenula wingei ; Inducible mutant ; Saccharomyces cerevisiae ; Saccharomyces kluyveri ; Sexual agglutinability ; Shmoo ; Synthetic analogues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three analogues of the peptidyl pheromone, α pheromone of Saccharomyces kluyveri, synthesized based on the amino acid sequence proposed by Sato et al. (Agric Biol Chem 45:1531–1533, 1981) were tested for both shmoo-inducing and agglutinability-inducing actions. Purified natural α pheromone of the yeast showed the highest activity among the peptides tested. When methionine in the peptides was oxidized, the activity decreased significatly. α Pheromone of S. kluyveri induced sexual agglutinability in a cells of Saccharomyces cerevisiae, and shmoo in a cells of S. cerevisiae and S. kluyveri. a Pheromone of S. kluyveri had no agglutinability-inducing action on α cells of S. cerevisiae. a Cells of S. kluyveri inactivated only α pheromone of the same species, but a cells of S. cerevisiae inactivated α pheromones of both S. cerevisiae and S. kluyveri.
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    Anaerobic growth of Saccharomyces cerevisiae in the absence of oleic acid and ergosterol? (1983)
    Springer
    Archives of microbiology 134 (1983), S. 64-67 
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    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.
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    Comparison of the killer toxin of several yeasts and the purification of a toxin of type K2 (1984)
    Springer
    Archives of microbiology 137 (1984), S. 357-361 
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    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Killer toxin ; Extracellular glycoprotein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A total of 13 killer toxin producing strains belonging to the genera Saccharomyces, Candida and Pichia were tested against each other and against a sensitive yeast strain. Based on the activity of the toxins 4 different toxins of Saccharomyces cerevisiae, 2 different toxins of Pichia and one toxin of Candida were recognized. The culture filtrate of Pichia and Candida showed a much smaller activity than the strains of Saccharomyces. Extracellular killer toxins of 3 types of Saccharomyces were concentrated and partially purified. The pH optimum and the isoelectric point were determined. The killer toxins of S. cerevisiae strain NCYC 738, strain 399 and strain 28 were glycoproteins and had a molecular weight of Mr=16,000. The amino acid composition of the toxin type K2 of S. cerevisiae strain 399 was determined and compared with the composition of two other toxins.
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    An examination of factors affecting the instability of Saccharomyces cerevisiae glucan synthetase in cell free extracts (1984)
    Springer
    Archives of microbiology 137 (1984), S. 209-214 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Glucan synthetase ; EDTA ; Magnesium ; Sucrose ; Fluoride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Yeast β(1–3) glucan synthetase is stimulated and stabilized by EDTA. Sucrose protects the enzyme from selfinactivaton. Preincubation of cell free extracts at low sucrose concentrations indicates a slow transition of the enzyme towards dissociation. Transition kinetics at 30° C and 0° C in the presence and in the absence of sucrose are interpreted assuming that a subunit is thermolabile in the free state and that sucrose increases its stability. Magnesium is deletereous for glucan synthetase in cell-free extracts. Chaotropic agents inactivate glucan synthetase according to their capacity to solubilize and depolymerize biological compounds. Fluoride plays a special role in the activation of glucan synthetase. Its action appears to be dependent on the presence of GTP (or other nucleotides). The role of all these agents on the activity and stability of the enzyme is interpreted in a unified scheme.
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    Selective inhibition of transition from sexual agglutination to zygote formation by ethyl N-phenylcarbamate in Saccharomyces cerevisiae (1984)
    Springer
    Archives of microbiology 137 (1984), S. 188-193 
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    ISSN: 1432-072X
    Keywords: Agglutination substance ; α Pheromone ; Cell cycle ; Ethyl N-phenylcarbamate ; Mating reaction ; Microtubules ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effects of ethyl N-phenylcarbamate (EPC) on the mating reaction of Saccharomyces cerevisiae were studied, with special attention on the effect on the α pheromone action. EPC inhibited zygote formation at a concentration which promoted induction of sexual agglutinability. EPC enhanced agglutinability induction by α pheromone, but inhibited α-pheromone-induced formation of large pearshaped cells in a mating type. The enhancement of agglutinability induction was accompanied with increased production of a agglutination substance and inhibition of α pheromone inactivation. EPC arrested the cell cycle of a cells probably in the step controlled by CDC19, CDC35, cAMP etc., just before the step controlled by CDC28, α pheromone etc.
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    Phosphorus-31 nuclear magnetic resonance studies of intracellular pH, phosphate compartmentation and phosphate transport in yeasts (1982)
    Springer
    Archives of microbiology 133 (1982), S. 83-89 
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    ISSN: 1432-072X
    Keywords: Candida utilis ; Saccharomyces cerevisiae ; Zygosaccharomyces bailii ; Compartmentation ; Vacuoles ; Internal pH ; Phosphate ; Glycolysis ; Nuclear magnetic resonance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 31P NMR spectra were obtained from suspensions of Candida utilis, Saccharomyces cerevisiae and Zygosaccharomyces bailii grown aerobically on glucose. Direct introduction of substrate into the cell suspension, without interruption of the measurements, revealed rapid changes in pH upon addition of the energy source. All 31P NMR spectra of the yeasts studied indicated the presence of two major intracellular inorganic phosphate pools at different pH environments. The pool at the higher pH was assigned to cytoplasmic phosphate from its response to glucose addition and iodoacetate inhibition of glycolysis. After addition of substrate the pH in the compartment containing the second phosphate pool decreased. A parallel response was observed for a significant fraction of the terminal and penultimate phosphates of the polyphosphate observed by 31P NMR. This suggested that the inorganic phosphate fraction at the lower pH and the polyphosphates originated from the same intracellular compartment, most probably the vacuole. In this vacuolar compartment, pH is sensitive to metabolic conditions. In the presence of energy source a pH gradient as large as 0.8 to 1.5 units could be generated across the vacuolar membrane. Under certain conditions net transport of inorganic phosphate across the vacuolar membrane was observed during glycolysis: to the cytoplasm when the cytoplasmic phosphate concentration had become very low due to sugar phosphorylation, and into the vacuole when the former concentration had become high again after glucose exhaustion.
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    Isolation, and biochemical and biological characterization of an a-mating-type-specific glycoprotein responsible for sexual agglutination from the cytoplasm of a-cells, in the yeast Saccharomyces cerevisiae (1984)
    Springer
    Archives of microbiology 140 (1984), S. 113-119 
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    ISSN: 1432-072X
    Keywords: Agglutination substance ; Cell-cell recognition ; Glycoprotein ; Mating ; Saccharomyces cerevisiae ; Sexual agglutinability ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An a-mating-type-specific substance responsible for sexual agglutination was purified to 397-times in specific activity (units/mg protein) from the cytoplasm of a-mating type cells. The purified substance gave a single band stained with PAS reagent but not with both Coomassie brilliant blue and silver staining reagent by polyacrylamide gel electrophoresis in the presence of 8 M urea. However, incorporation of [35S]methionine and Lowry reaction clearly indicate that the substance is a glycoprotein. The substance specifically masked sexual agglutinability of cells of the opposite mating type α, indicating univalent action. The substance is a glycoprotein with a carbohydrate content of 90%, a pI of 4.5, and a molecular weight of 130,000. The substance was inactivated by 2-mercaptoethanol and proteolytic enzymes but not by glycolytic enzymes. The substance formed a complementary complex having no biological activity when mixed with α-agglutination substance from the wall or cytoplasm of α-cells in vitro.
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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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    Effect of ozone on ATP, cytosolic enzymes and permeability of Saccharomyces cerevisiae (1987)
    Springer
    Archives of microbiology 147 (1987), S. 105-108 
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    ISSN: 1432-072X
    Keywords: Ozone ; Yeast ; Saccharomyces cerevisiae ; ATP ; Nucleotides ; Permeability ; Cytosolic enzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Treatment of a yeast suspension with ozone inactivates a number of cytosolic enzymes. Among 15 studied, the most drastic inactivation was found for glyceraldehyde-3-phosphate dehydrogenase and to lesser extents: NAD-glutamate dehydrogenase, pyruvate decarboxylase, phosphofructokinase-1 and NAD-alcohol dehydrogenase. Ozone treatment also effects the quantity of ATP and of other nucleoside triphosphates, reducing to about 50% of the initial value. The ATP missing in the cells appears in the medium. NAD and protein also accumulate in the medium suggesting that the yeast cells have been permeabilized. Permeabilization of the yeast cells by treatment with ozone preceeds the inactivation of glyceraldehyde-3-phosphate dehydrogenase and other cytosolic enzymes.
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    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 
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    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating reaction ; Zygote formation ; Mating pheromone ; Fatty acid ; Arachidonic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effect of exogenous fatty acids on zygote formation in Saccharomyces cerevisiae was studied. Arachidonic and oleic acids considerably stimulated zygote formation, but other fatty acids tested, linoleic, linolenic, stearic and palmitic acids, did not. Pretreatment experiments with arachidonic acid showed that the stimulation of zygote formation by the fatty acid required the presence of mating pheromone.
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    Isolation and composition of the constitutive agglutinins from haploid Saccharomyces cerevisiae cells (1987)
    Springer
    Archives of microbiology 148 (1987), S. 208-212 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Yeast mating ; Cell-cell recognition ; Sexual agglutination ; Agglutinins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sex-specific agglutinins from the cell surface of haploid cells of Saccharomyces cerevisiae (X2180, mta and mtα) were purified and analysed. The constitutive agglutinin from mta cells was extractable with 3 mM dithiothreitol. It was shown to be a glycoprotein (3% mannose) with an apparent Mr of 43,000 based on gel filtration, but in SDS-PAGE it behaved as a much smaller molecule (Mr between 18,000 and 26,000). About one in three amino acids was a hydroxyamino acid. Its biological activity was resistant to boiling for 1 h, but sensitive to pronase. Intact mtα cells retained their agglutinability in the presence of dithiothreitol but limited trypsinizing released a biologically active agglutinin fragment. It had an apparent Mr of 320,000 (gel filtration). When analysed by SDS-PAGE, a single diffuse band with an apparent Mr of 225,000 was observed. The protein was 94% (w/w) mannose with a trace of N-acetyl glucosamine. Its biological activity was almost completely lost after boiling for 1 h. Both agglutinins behaved as monovalent molecules and specifically inhibited the biological activity of both noninduced and pheromone-induced cells. Pheromone treatment of mta cells resulted in an apparent 32-fold increase in agglutinin activity at the cell surface, whereas pheromone treatment of mtα cells only doubled the apparent agglutinin activity.
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    Chloroquine, a lysosomotropic agent, inhibits zygote formation in yeast (1988)
    Springer
    Archives of microbiology 151 (1988), S. 20-25 
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    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating ; Zygote formation ; Chloroquine ; Lysosomotropic agent ; Plasma membrane ; Cell fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Haploid cells of opposite mating type of Saccharomyces cerevisiae conjugate to form zygote. During the conjugation process, the degradation or reorganization of the cell wall and the fusion of the two plasma membranes take place. Since chloroquine inhibits cellular events associated with the reorganization of the plasma membrane, the effect of the drug on conjugation was studied. Chloroquine at a concentration, at which cell growth was not retarded, inhibited zygote formation, while it did not affect other mating functions, such as sexual agglutination, production of and response to mating pheromone. Cells in a mating culture containing chloroquine formed no “prezygote” suggesting that they were not prepared for entering into fusion process. The inhibitory effect of chloroquine was reversible as cells formed zygote when they were washed after treatment with chloroquine. Zygote formation was unaffected in cells possessing chlorquine within vacuoles after incubation with the drug in complete medium (YPD) at pH 7.5, followed by washing. This suggests that chloroquine inhibits zygote formaton by adsorbing to the plasma membrane of S. cerevisiae.
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    Effects of nucleotides and divalent cations on phospholipase activity in Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 154-158 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Yeast ; Phospholipase B ; Lysophospholipase ; Enzyme inhibition ; AMP ; Unesterified fatty acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Divalent cations activate the lysophospholipase and transacylase reactions catalyzed by the same enzymes in the yeast Saccharomyces cerevisiae. The activation was observed at neutral pH, but not at the pH optimum of lysophospholipase/transacylase, near 3.5. Adenine nucleotides, especially AMP and ADP, are strong inhibitors of the same group of enzymes. Half maximal inhibition by AMP was found at a concentration of about 20 μM. The inhibition by nucleotides in low concentrations is enhanced by divalent cations.
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    The glucan-chitin complex in Saccharomyces cerevisiae (1981)
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    Archives of microbiology 130 (1981), S. 312-318 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Bud scar ; Scar ring ; Glucan ; Chitin ; Mannan ; Cytology ; Electron and X-ray diffractions ; Physico-chemical characterization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Scar rings (SR) from scarless cells at the early stages of budding and mature bud scars from Saccharomyces cerevisiae isolated by both chemical and enzymic treatment of cell walls were observed by selected-area electron diffraction (SAED), X-ray diffraction and electron microscopy with simultaneous physico-chemical characterization (including molar mass, intrinsic viscosity and crystallite size) of α-chitin and glucan. The SR, composed of glucan with strong 0.608; 0.397 and 0.293 nm X-ray reflections, was formed at the start of budding. The SAED patterns of α-chitin both in the adjacent circular zone and in the parts of newly formed primary septum (PS), observed when the development of the PS started, did not differ from those of α-chitin in the single mature bud scar. The bud scar consisted of α-chitin, glucan and mannan and their content, as well as the crystallite size of chitin, depended on the mode of preparation.
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    Localization of trehalase in vacuoles and of trehalose in the cytosol of yeast (Saccharomyces cerevisiae) (1982)
    Springer
    Archives of microbiology 131 (1982), S. 298-301 
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    ISSN: 1432-072X
    Keywords: Yeast ; Protoplast ; Compartmentation ; Vacuole ; Trehalose ; Trehalase ; Carbohydrate metabolism ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of Saccharomyces cerevisiae synthesized and degraded trehalose when they were incubated in a medium containing traces of glucose and acetate. Such protoplasts were gently lyzed by the polybase method and a particulate and soluble fraction was prepared. Trehalose was found in the soluble fraction and the trehalase activity mostly in the particulate fraction which also contained the vacuoles besides other cell organelles. Upon purification of the vacuoles, by density gradient centrifugation, the specific activity of trehalase increased parallel to the specific content of vacuolar markers. This indicates that trehalose is located in the cytosol and trehalase in the vacuole. It is suggested that trehalose, in addition to its role as a reserve may also function as a protective agent to maintain the cytosolic structure under conditions of stress.
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    Inhibition of yeast tRNATrp aminoacylation by 4-methyltryptophan (1981)
    Springer
    Archives of microbiology 129 (1981), S. 146-149 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Inhibition of tryptophanyl-tRNA synthetase ; Mode of action of tryptophan analogues ; Tryptophan analogue degradation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of the tryptophan analogue 4-methyltryptophan in Saccharomyces cerevisiae has been investigated. 4-Methyltryptophan inhibits the aminoacylation of tryptophan specific transfer ribonucleic acid (tRNATrp). The mode of inhibition is competitive and the analogue is not charged onto tRNATrp. Thus 4-methyltryptophan application depletes the cells from charged tRNATrp. As a consequence cell growth and protein synthesis are strongly reduced. 4-Methyltryptophan is degraded efficiently in culture media inoculated with the wild type strain; the effects of 4-methyltryptophan were therefore found to be transient.
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    Sex pheromones of the yeast Hansenula wingei and their relationship to sex pheromones in Saccharomyces cerevisiae and Saccharomyces kluyveri (1981)
    Springer
    Archives of microbiology 129 (1981), S. 281-284 
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    ISSN: 1432-072X
    Keywords: Hansenula wingei ; Induction ; Saccharomyces cerevisiae ; Saccharomyces kluyveri ; Sex pheromone ; Sexual agglutinability ; Sexual agglutination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The yeast, Hansenula wingei has two mating types designated 5 and 21. Cells of each mating type were found to produce mating type-specific sex pheromone which induces sexual agglutinability of the opposite mating type. Crude fractions of these pheromones were prepared by using an Amberlite CG 50 (H+ type) column. The agglutinability-inducing action of the pheromones required glucose as carbon source, but no external nitrogen source. The action of the pheromones was inhibited by 5 μg/ml cycloheximide. The optimum pH for the pheromone action was 4.0. α Pheromones of Saccharomyces cerevisiae and Saccharomyces kluyveri induced sexual agglutinability of 5 mating type cells but did not that of 21 mating type cells. a Pheromones of the Saccharomyces yeasts had no effect on both 5 and 21 mating type cells. The sex pheromones of H. wingei had no effect on the sexual agglutinability of inducible a cells of S. cerevisiae. From the experimental results obtained so far, we propose to call 5 and 21 mating types in H. wingei a and α mating types, respectively.
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    Tryptophan degradation in Saccharomyces cerevisiae: Characterization of two aromatic aminotransferases (1982)
    Springer
    Archives of microbiology 133 (1982), S. 242-248 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Tryptophan degradation to tryptophol ; Degradation-defective mutant strain ; Aromatic aminotransferases ; Tryptophan accumulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tryptophan was found to be degraded in Saccharomyces cerevisiae mainly to tryptophol. Upon chromatography on DEAE-cellulose two aminotransferases were identified: Aromatic aminotransferase I was constitutively synthesized and was active in vitro with tryptophan, phenylalanine or tyrosine as amino donors and pyruvate, phenylpyruvate or 2-oxoglutarate as amino acceptors. The enzyme was six times less active with and had a twenty times lower affinity for tryptophan (K m=6 mM) than phenylalanine or tyrosine. It was postulated thus that aromatic aminotransferase I is involved in vivo in the last step of tyrosine and phenylalanine biosynthesis. Aromatic aminotransferase II was inducible with tryptophan but also with the other two aromatic amino acids either alone or in combinations. With tryptophan as amino donor the enzyme was most active with phenylpyruvate and not active with 2-oxoglutarate as amino acceptor; its affinity for tryptophan was similar as for the other aromatic amino acids (K m=0.2–0.4 mM). Aromatic aminotransferase II was postulated to be involved in vivo mainly in the degradation of tryptophan, but may play also a role in the degradation of the other aromatic amino acids. A mutant strain defective in the aromatic aminotransferase II (aat2) was isolated and its influence on tryptophan accumulation and pool was studied. In combination with mutations trp2 fbr, aro7 and cdr1-1, mutation aat2 led to a threefold increase of the tryptophan pool as compared to a strain with an intact aromatic aminotransferase II.
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    A potassium transport mutant of Saccharomyces cerevisiae (1985)
    Springer
    Archives of microbiology 143 (1985), S. 88-93 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Potassium transport mutant ; Rubidium transport ; Sodium transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mutant in Saccharomyces cerevisiae required one hundred times more K+ than wild type for the same half maximal growth rate. Mutant cells and wild type cells grown at millimolar K+ did not show significant differences in Rb+ transport. In the mutant, a rapid K+ loss induced by azide or incubation (4 h) in K+-free medium decreased the Rb+ transport K m by one half; in the wild type, those treatments decreased the Rb+ K m twenty and one hundred times, respectively. Mutant and wild type did not show significant differences in Na+ transport and in the Na+ inhibition of Rb+ transport, either in normal-K+ cells or in K+-starved cells. The results suggest that either two systems or one system with two interacting sites mediate K+ transport in S. cerevisiae.
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    Regulation of β-exoglucanase activity production by Saccharomyces cerevisiae in batch and continuous culture (1985)
    Springer
    Archives of microbiology 143 (1985), S. 143-146 
    Details
    ISSN: 1432-072X
    Keywords: Exoglucanase ; Saccharomyces cerevisiae ; Continuous culture ; Growth rate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The rate of synthesis and secretion of exo-1–3-β-glucanase activity closely paralleled the specific rate of growth in exponentially growing Saccharomyces cerevisiae cells in batch culture. When the stationary phase was reached both synthesis and secretion stopped. No activity was synthesized when the cells were maintained in carbon sources that did not allow them to grow. Studies in continuous culture indicate a strong relationship between the synthesis of exoglucanase activity and the specific growth rate. These results are taken as evidence of an essential role of this activity during the yeast budding cycle.
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    Arrangement of genes TRP1 and TRP3 of Saccharomyces cerevisiae strains (1985)
    Springer
    Archives of microbiology 142 (1985), S. 383-388 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Tryptophan biosynthesis ; General control ; Systematic by Southern hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The tryptophan biosynthetic genes TRP1 and TRP3 and partly also TRP2 and TRP4 have been compared by the technique of Southern hybridization and enzyme measurements in twelve wild isolates of Saccharomyces cerevisiae from natural sources of different continents, in the commonly used laboratory strain S. cerevisiae X2180-1A and in a Kluyveromyces marxianus strain. We could classify these strains into four groups, which did not correlate with their geographical distribution. In no case are the TRP3 and TRP1 genes fused as has been found in other ascomycetes. Two strains were found which, in contrast to strain X2180-1A, show derepression of gene TRP1. Two examples are discussed to demonstrate the usefulness of Southern hybridizations for the identification of closely related strains.
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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)
    Springer
    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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    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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    Genetic analysis of inducible sexual agglutination ability in the yeast Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 198-202 
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    ISSN: 1432-072X
    Keywords: Sexual agglutination ; Mating ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic regulation of the inducibility of sexual agglutination ability in the yeast Saccharomyces cerevisiae was studied. Detailed analysis of the degree of sexual agglutination was carried out; it showed that a greater number of genes are involved in the regulation of inducible sexual agglutination in strain H1-0 than previously assumed. Although dominancy of inducible phenotype over constitutive was confirmed, the effectiveness of one gene changing the constitutive phenotype to the inducible seemed to be somewhat low. Quantity per cell of agglutination substances responsible for sexual agglutination increased as the agglutination ability became greater.
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    A similar protein portion for two exoglucanases secreted by Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 391-398 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Exoglucanases ; Purification ; Protein moieties ; Tunicamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Exoglucanase (exo-1,3-β-D-glucan glycohydrolase, EC 3.2.1.56) activity secreted by Saccharomyces cerevisiae into the culture medium was separated by ion exchange chromatography into two glycoprotein isoenzymes which contributed 10% (exoglucanase I) and 90% (exoglucanase II) towards the total activity. Analysis of the “in vitro” deglycosylated products by polyacrylamide gel electrophoresis under native or denaturing conditions indicated that the protein portions of both exoglucanases exhibited identical mobility, each one consisting of two polypeptides with M r of 47000 and 48000. The same profile was shown by the exoglucanase secreted in the presence of tunicamycin. Antibodies raised against the protein portion of exoglucanase II did react with both native exoglucanases and their deglycosylated products with a pattern indicative of immunological identity. Digestion of the “in vitro” deglycosylated products of both exoglucanases with Staphylococcus aureus V-8 protease or trypsin generated the same proteolytic fragments in each case. Only exoglucanase II was secreted by protoplasts. These and previously reported results indicate that the protein portions of both isoenzymes may be the product of the same gene (or a family of related genes), and that exoglucanase I is a product of enzyme II, modified by a process occurring beyond the permeability barrier of the cell.
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    Nucleotide modification of tRNA in. the yeast Saccharomyces cerevisiae is not Affected by the ψ factor which modulates suppression efficiency (1981)
    Springer
    Current genetics 3 (1981), S. 163-165 
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    ISSN: 1432-0983
    Keywords: Suppressors-tRNA ; Saccharomyces cerevisiae ; Nucleotide modification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have examined the tRNAs of two related strains of Saccharomyces cerevisiae, ψ + and ψ −, which differ with respect to an extrachromosomal genetic element that modulates the expression of genotypic and phenotypic suppression. Both the pattern of tRNAs synthesized and the level of nucleotide modification of several selected tRNA species were found to be the same in the ψ + and ψ − strains.
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    Construction of new yeast vectors and cloning of the nif (nitrogen fixation) gene cluster of Klebsiella pneumoniae in yeast (1981)
    Springer
    Current genetics 3 (1981), S. 173-180 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Yeast vectors ; Cosmids ; nif genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two vectors, termed pG63.11 (7.6 Kb) and pHCG3 (9.6 Kb), suitable for yeast transformation have been constructed. The pHCG3 vector has cosmid properties. Both vectors contain a single 3.3 Kb EcoRI-HindIII fragment of yeast origin which carries the yeast URA3 gene (1.1 Kb) and the origin of replication of the 2 µm plasmid (2.2 Kb). They confer ampicillin resistance and they contain 5 unique EcoRI,HpaI,HindIII,BamHI and SalI restriction sites. Cosmid pHCG3 was used to clone the nitrogen fixation (nif) gene cluster of Klebsiella pneumoniae carried by twoHindIII fragments of 17 and 26 Kb, respectively. The resulting cosmid, termed pGPC875 (53 Kb) which conferred a Nif+ phenotype to Escherichia coli, was introduced in yeast by transformation. No acetylene reduction activity was detectable in the transformants. However it was shown that the entire information for nitrogen fixation can be replicated and maintained intact in yeast for more than 50 generations of growth.
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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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    Cloning and integrative deletion of the RAD6 gene of Saccharomyces cerevisiae (1984)
    Springer
    Current genetics 8 (1984), S. 559-566 
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    ISSN: 1432-0983
    Keywords: DNA repair ; Saccharomyces cerevisiae ; Cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three overlapping plasmids were isolated from a YEp24 library, which restore Rad+ functions to rad6-1 and rad6-3 mutants. Different subclones were made and shown to integrate by homologous recombination at the RAD6 site on chromosome VII, thus verifying the cloned DNA segments to be the RAD6 gene and not a suppressor. The gene resides in a 1.15 kb fragment, which restores Rad+ levels of resistance to U.V., MMS and γ-rays to both rad6-1 and rad6-3 strains. It also restores sporulation ability to rad6-1 diploids. Integrative deletion of the RAD6 gene was shown not to be completely lethal to the yeast. Our results suggest that the RAD6 gene has some cell cycle-specific function(s), probably during late S phase.
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    Hybridization of Saccharomyces cerevisiae with Candida utilis through protoplast fusion (1984)
    Springer
    Current genetics 8 (1984), S. 575-580 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Candida utilis ; Protoplast fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Auxotrophic mutants of Saccharomyces cerevisiae and Candida utilis were hybridized through protoplast fusion. Spontaneous, UV- and FPA-induced mitotic segregation indicated that after cell fusion, exclusion of the S. cerevisiae nucleus or nuclear fusion followed by preferential loss of S. cerevisiae chromosomes can take place. Some of the hybrids were stable. One of them, expressed mating and sporulation functions of the S. cerevisiae parent. Thus, markers from both parents could be recovered as mitotic and meiotic segregants.
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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 
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    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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    An α-mating-type-specific mutation causing specific defect in sexual agglutinability in the yeast Saccharomyces cerevisiae (1985)
    Springer
    Current genetics 9 (1985), S. 185-189 
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    ISSN: 1432-0983
    Keywords: Agglutination substance ; Mating type ; Non-agglutinable mutant ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have identified a recessive α-mating-type-specific gene agαl causing agglutinability defect without significant effects on other sexual activities. a cells carrying agαl showed sexual agglutination with α cells but α cells carrying agαl showed sexual agglutination with neither α cells nor a cells. α cells carrying agαl produced α pheromone and responded to a pheromone just like wild α cells. α cells carrying agαl showed a little decreased but significant mating ability when tested on solid media or membrane filter. The agαl mutant is different from any α-specific ste mutants found so far in many sexual activities. The agαl gene is recessive, and unlinked to the mating type locus. Biological significance of the α mating type agglutinability is discussed based on the results obtained with the mutant.
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    Selection in yeast I: Assessing genetic stability and relative fitness of commercial yeasts (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 159-165 
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    ISSN: 1476-5535
    Keywords: Yeast ; Genetic stability ; Saccharomyces cerevisiae ; Selection ; Reproductive fitness
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The potential for changes in allele frequencies in yeast populations by selection was examined. Cells from the wine yeastSaccharomyces cerevisiae (strain Montrachet) were grown over a large number of generations using two different culturing techniques, each with two variations: serial transfers on WLN agar plates with and without UV irradiation, and continuous culture in autoclaved and in filter-sterilized grape must. A low frequency of variant isozyme patterns was found in samples taken at the end of the experiment. Growth rates in must and on agar plates were also examined, and it was found that all samples were faster-growing than the original strain, to varying degrees. Applications for the selection system developed are discussed.
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    Biotransformation of aromatic aldehydes bySaccharomyces cerevisiae: Investigation of reaction rates (1989)
    Springer
    Journal of industrial microbiology and biotechnology 4 (1989), S. 49-53 
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    ISSN: 1476-5535
    Keywords: l-Phenylacetyl carbinol ; Saccharomyces cerevisiae ; Yeast ; Benzaldehyde ; Biotransformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The rate of production ofl-phenylacetyl carbinol bySaccharomyces cerevisiae in reaction mixtures containing benzaldehyde with sucrose or pyruvate as cosubstrate was investigated in short 1 h incubations. The effect of yeast dose rate, sucrose and benzaldehyde concentration and pH on the rate of reaction was determined. Maximum biotransformation rates were obtained with concentrations of benzaldehyde, sucrose and yeast of 6 g, 40 g and 60 g/l, respectively. Negligible biotransformation rates were observed at a concentration of 8 g/l benzaldehyde. The reaction had a pH optimum of 4.0–4.5. Rates of bioconversion of benzaldehyde and selected substituted aromatic aldehydes using both sucrose and sodium pyruvate as cosubstrate were compared. The rate of aromatic alcohol production was much higher when sucrose was used rather than pyruvate.o-Tolualdehyde and 1-chlorobenzaldehyde were poor substrates for aromatic carbinol formation although the latter produced significant aromatic alcohol in sucrose-containing media. Yields of 2.74 and 3.80 g/l phenylacetyl carbinol were produced from sucrose and pyruvate, respectively, in a 1 h reaction period.
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    Selection in yeast II: The fitness distribution of new mutations inSaccharomyces cerevisiae and its use in a computer simulation (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 167-174 
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    ISSN: 1476-5535
    Keywords: Selection ; Yeast ; Fitness distribution ; Mutation ; Saccharomyces cerevisiae ; Computer simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The fitness distribution of new mutations inSaccharomyces cerevisiae strain Montrachet was determined for cells on agar irradiated for four periods of time with ultraviolet light. The fitness distributions were obtained by converting a large number of colony diameters into relative fitnesses. The distributions were then used to perform a computer simulation with the purpose of predicting the potential of a stock culture to increase in general fitness through selection, given a frequency and magnitude of mutations.
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    Optically monitoring Baker's yeast (Saccharomyces cerevisiae) growing in an air-fluidized/expanded potato starch matrix (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 187-193 
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    ISSN: 1476-5535
    Keywords: Saccharomyces cerevisiae ; Fermentation ; Air-fluidized fermentation ; Semi-solid fermentation ; Yeast cell concentration in starch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary In order to study cell behavior in solid fermentation processes, model systems using gelatin and starch have been developed to track Baker's yeast growth. The difficulty in estimating the cell concentration within solid materials arises because both the solid material and the cellular material contribute to the measurement (such as optical resistance). In general, however, the two materials cannot be easily separated, hence the need to measure the cells along with the solid supporting material. A simple spectrophotometric method has previously been shown to work well in both aerated submerged batch cultures and aerated static solid cultures. The optical approach is applied here to monitor a more complex solidified system: cell growth in a novel air-fluidized/expanded bed of yeast growing on a starch matrix. Conventional assays for reducing sugar, total extracellular protein, and extracellular lysine were also applied to monitor yeast behavior in this new system.
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    Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation (1988)
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    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
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    ISSN: 1476-5535
    Keywords: Osmotic pressure ; Intracellular ethanol ; Yeast ; Nutrient ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The intracellular accumulation of ethanol in yeast and its potential effects on growth and fermentation have been topics of controversy for the past several years. The determination of intracellular ethanol based on the exclusion of [14C]sorbitol to estimate aqueous cell volume was used to examine the question of intracellular ethanol accumulation. An intracellular accumulation of ethanol inSaccharomyces cerevisiae was observed during the early stages of fermentation. However, as fermentation continued, the intracellular and extracellular concentrations of ethanol became similar. Increasing the osmotic pressure of the medium with glucose or sorbitol was observed to cause an increase in the intracellular ethanol concentration. Associated with this was a decrease in yeast growth and fermentation rates. In addition, increasing the osmotic pressure of the medium was observed to cause an increase in glycerol production. Supplementation of the media with excess peptone, yeast extract, magnesium sulfate and potassium phosphate was found to relieve the detrimental effects of high osmotic pressure. Under these conditions, though, no effect on the intracellular and extracellular ethanol distribution was observed. These results indicate that nutrient limitation, and not necessarily intracellular ethanol accumulation, plays a key role during yeast fermentations in media of high osmolarity.
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    The reconstitution of oxidative phosphorylation in mitochondria isolated from a ubiquinone-deficient mutant ofSaccharomyces cerevisiae (1982)
    Springer
    Journal of bioenergetics and biomembranes 14 (1982), S. 159-169 
    Details
    ISSN: 1573-6881
    Keywords: Respiratory chain ; ATP synthesis ; mitochondria ; ubiquinone ; Saccharomyces cerevisiae ; cytochrome oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Mitochondria, isolated from the ubiquinone-deficient nuclear mutant ofSaccharomyces cerevisiae E3-24, are practically unable to oxidize exogenous substrates. Respiratory activity, coupled to ATP synthesis, can, however, be reconstituted by the simple addition of ethanolic solutions of ubiquinones. A minimal length of the isoprenoid side chain (≥3) was required for the restoration. Saturation of the reconstitution required a large amount of exogeneous ubiquinone, in excess over the normal content present in the mitochondria of the wild type strain. A similar pattern of reconstituted activities could be also obtained using sonicated inverted particles. Mitochondria and sonicated particles are also able to carry out a dye-mediated electron flow coupled to ATP synthesis in the absence of added ubiquinone, using ascorbate or succinate as electron donor. This demonstrates that the energy conserving mechanism at the third coupling site of the respiratory chain is fully independent of the presence of the large mobile pool of ubiquinone in the membrane.
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    Biogenesis of mitochondria: Defective yeast H+-ATPase assembled in the absence of mitochondrial protein synthesis is membrane associated (1984)
    Springer
    Journal of bioenergetics and biomembranes 16 (1984), S. 561-581 
    Details
    ISSN: 1573-6881
    Keywords: H+-ATPase complex ; assembly defect ; Saccharomyces cerevisiae ; mitochondrial biogenesis ; membrane association
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract We have investigated the extent to which the assembly of the cytoplasmically synthesized subunits of the H+-ATPase can proceed in a mtDNA-less (rho°) strain of yeast, which is not capable of mitochondrial protein synthesis. Three of the membrane sector proteins of the yeast H+-ATPase are synthesized in the mitochondria, and it is important to determine whether the presence of these subunits is essential for the assembly of the imported subunits to the inner mitochondrial membrane. A monoclonal antibody against the cytoplasmically synthesized β-subunit of the H+-ATPase was used to immunoprecipitate the assembled subunits of the enzyme complex. Our results indicate that the imported subunits of the H+-ATPase can be assembled in this mutant, into a defective complex which could be shown to be associated with the mitochondrial membrane by the analysis of the Arrhenius kinetics of the mutant mitochondrial ATPase activity.
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    Terminal segment of Kluyveromyces lactis linear DNA plasmid pGKL2 supports autonomou sreplication of hybrid plasmids in Saccharomyces cerevisiae (1987)
    Springer
    Current genetics 12 (1987), S. 99-104 
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    ISSN: 1432-0983
    Keywords: ARS ; Linear DNA killer plasmid ; Replication ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By use of linear DNA plasmid pGKL2 from the yeast Kluyveromyces lactis we have constructed hybrid plasmids carrying a LEU2 gene of Saccharomyces cerevisiae as a selectable marker. The replication properties of hybrid plasmids in yeasts were investigated. We demonstrated that the insertion of a LEU2 gene into pGKL2 resulted in circularization of the hybrid plasmids and pGKL2 segment supported autonomous replication of the plasmids. Moreover, the hybrid plasmids propagated autonomously, independently of the presence of the natural pGKL2 plasmid.
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    Construction of brewer's yeasts secreting fungal endo-ß-glucanase (1987)
    Springer
    Current genetics 12 (1987), S. 413-420 
    Details
    ISSN: 1432-0983
    Keywords: Glucanolytic brewer's yeast ; Endo-β-1,4-glucanase ; Chromosomal integration ; Transformation ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Barley β-glucans present in wort reduce beer filtrability and cause hazes and precipitates in the finished beer. The endo-β-1,4-glucanase enzyme, EGI, found in the filamentous fungus Trichoderma reesei, is capable of efficiently hydrolyzing these β-glucans. The cDNA copy of the eg11 gene, which codes for the EGI enzyme, was coupled to yeast regulatory sequences and transferred to a brewer's yeast using the yeast copper chelatin gene CUP1 as a selection marker in the transformation. The eg11 gene was transferred to the yeast both on a multicopy plasmid and on an integrating plasmid. In both cases, highly glycosylated, active EGI enzyme was secreted into the medium. Barley β-glucans present in wort were efficiently hydrolyzed by the recombinant brewer's yeast.
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    Nuclear mutations affecting the stability of the mitochondrial genome inS. cerevisiae (1985)
    Springer
    Current genetics 10 (1985), S. 171-177 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Nuclear mutations ; Mitochondrial DNA stability ; Uncoupling of phenotypes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have studied a pleiotropic mutationpetD inS. cerevisiae which both confers the inability to grow on glycerol (Gly−) and greatly increases the frequency of cytoplasmic petites (Het). The first phenotype, Gly−, is recessive, whereas the second, Het, is dominant. Genetic and biochemical analysis showed that the majority of the petites inpetD strains are not of therho° type (completely lacking mit-DNA),but of therho − type (containing partially deleted mit-DNA). This finding and the fact that the phenotype Het is dominant argue in favour of the involvement of thepetD product in the excision process of the mit-DNA. Another nuclear mutation,mod, was shown to exhibit a dominant epistasy with respect to the Het phenotype of the mutationpetD. Two types of Gly+ revertants frompetD mutants were isolated:rpa revertants, which restore completely the wild-type phenotype, andrpb revertants, which restore only the growth on glycerol, but still allow the production of high frequencies of cytoplasmic petites. Thus the mutationsmod andrpb permit the genetic uncoupling of two phenotypes induced by the mutationpetD.
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    Saccharomyces cerevisiae strains sensitive to inorganic mercury (1985)
    Springer
    Current genetics 10 (1985), S. 179-185 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Resistance ; Mercury ; Tyrosine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary From a cross of two strains ofSaccharomyces cerevisiae, both of which had the same (wild type or normal) level of resistance to inorganic mercury, segregants having three distinguishable resistance levels, normal, sensitive and semi-sensitive, were obtained. Genetic analyses of the parents and the progeny indicated that the levels of inorganic mercury sensitivity were determined by three distinct loci,HGS1, HGS2 andMSM1. The recessive allele of theHGS1 locus,hgsl-1, and the codominant allele of theHGS2 locus,HGS2-1, were necessary for the sensitive phenotypes, and alleles in theMSM1 locus,MSMI-1 andmsml-2, were responsible for the different sensitivity levels. In short, the strains of genotypeshgs1-1 HGS2-1 msml-2 andhgsl-1 HGS2-1 MSMI-1 were sensitive and semi-sensitive, respectively, while the strains of all other genotypes were normal. Although thehgs1-1 allele was identified as thearo7-1 mutation which confers deficiency of tyrosine and phenylalanine, mutations such asaro1B (deficiency of tyrosine, phenylalanine and tryptophan) andtyr1 (deficiency of tyrosine) had similar effects asaro7-1 on inorganic mercury sensitivity. From these results we conclude that theHGS2-1 allele causes inorganic mercury sensitivity when the cells are defective in the tyrosine biosynthesis. In fact, addition of tyrosine to the growth medium containing inorganic mercury resulted in increase of colony forming ability of the sensitive strains.
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    Saccharomyces cerevisiae strains sensitive to inorganic mercury (1985)
    Springer
    Current genetics 10 (1985), S. 187-195 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Inorganic mercury ; Catabolite regulation ; Sugar uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Saccharomyces cerevisiae strains sensitive to inorganic mercury (Ono and Sakamoto 1985) did not grow well on the medium rich in glucose and poor in peptone. This growth inhibition, like growth inhibition caused by inorganic mercury, was relieved by exogenous tyrosine. Sugars such as fructose and mannose were as inhibitory as glucose, but glycerol was not at all. Galactose was inhibitory but not so much as glucose. Agal2l mutation (defective in galactose uptake) partly relieved growth inhibition caused by excess galactose. Moreover, it was found that some of revertants which gained ability to grow well in the presence of excess glucose were defective in the glucose uptake. From these observations, we conclude that growth inhibition of the inorganic mercury sensitive strains by excess sugar is a consequence of the catabolite regulation. In other words, the inorganic mercury sensitive strains are hyper-sensitive to the catabolite regulation due to the presence of theHGS2-1 allele.
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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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    Chromosomal mapping of the uracil permease gene of Saccharomyces cerevisiae (1986)
    Springer
    Current genetics 11 (1986), S. 93-96 
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    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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    Phenotypic differences between induced and spontaneous 2μ-plasmid-free segregants of Saccharomyces cerevisiae (1987)
    Springer
    Current genetics 11 (1987), S. 415-418 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; 2 μ plasmids ; Plasmid free segregants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The maximum specific growth rates (μmax) of 2 μ-plasmid-free ([cir°]) segregants of three haploid and one diploid strain of Saccharomyces cerevisiae have been determined and compared with the μmax of their 2 μ-plasmid-containing ([cir +]) progenitors. Two classes of [cir°] strains have been examined: those induced by transformation with a 2 μ-based recombinant plasmid according to the method of Dobson et al. (1980) and those isolated as spontaneous [cir°] segregants from glucose-limited continuous cultures. The μmax of the spontaneous [cir°] segregants was not found to differ significantly from that of their [cir +] parents. In all cases, however, the induced [cir°] strains had a μmax which was significantly less than that of their [cir +] counterparts. This effect was particularly marked in the case of the diploid strain where a 34% reduction in μmax was observed. The implications of these results are discussed in terms of the effect of the transformation process on host yeast cells.
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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 
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    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.
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    Comparison of Tetrahymena ARS sequence function in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe (1988)
    Springer
    Current genetics 14 (1988), S. 225-233 
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    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.
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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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    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 
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    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.
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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 
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    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.
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    The glucose- and ethanol-dependent regulation of PDC1 from Saccharomyces cerevisiae are controlled by two distinct promoter regions (1988)
    Springer
    Current genetics 14 (1988), S. 337-344 
    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.
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    Molecular cloning of the ARG7 gene of Schizosaccharomyces pombe encoding argininosuccinate lyase (1988)
    Springer
    Current genetics 14 (1988), S. 381-385 
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    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.
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    A galactose-dependent cmd1 mutant of Saccharomyces cerevisiae: involvement of calmodulin in nuclear division (1989)
    Springer
    Current genetics 15 (1989), S. 113-120 
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    ISSN: 1432-0983
    Keywords: Calmodulin mutant ; Nuclear division ; Chromosome stability ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The coding region of a yeast calmodulin gene was fused to a galactose-inducible GAL1 promoter, and a conditional-lethal mutant of Saccharomyces cerevisiae, in which the expression of calmodulin was regulated by galactose, was constructed. The mutant grew normally in galactose medium, but in glucose medium, in which the promoter was repressed, it ceased growing after 12–15 h. The growth arrest was associated with a decrease in intracellular calmodulin levels: after 12h, no intracellular calmodulin protein was detectable. Analysis of the terminal phenotype showed that when the cell stopped growing, it had a bud, a nucleus after S-phase and a short mitotic spindle. Thus, the defect was mainly in nuclear division. Bud growth was partially inhibited in these cells: 27% of the cells stopped growing with a small bud. Furthermore, calmodulin-deficient cells showed elevated rates of chromosome loss, possibly as the result of a defect in the precise segregation of chromosomes.
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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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    The allosuppressor gene SAL4 encodes a protein important for maintaining translational fidelity in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 537-543 
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    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.
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    Generation of an ilv bradytrophic phenocopy in yeast by antisense RNA (1988)
    Springer
    Current genetics 13 (1988), S. 283-289 
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    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.
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    The Saccharomyces cerevisiae RAD2 gene complements a Schizosaccharomyces pombe repair mutation (1989)
    Springer
    Current genetics 15 (1989), S. 27-30 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Repair ; Complementation ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two Saccharomyces cerevisiae genes necessary for excision repair of UV damage in DNA, RAD1 and RAD2, were introduced individually, on a yeast shuttle vector, into seven Schizosaccharomyces pombe mutants — rads1, 2, 5, 13, 15,16 and 17. The presence of the cloned RAD1 gene did not affect survival of any of the S. pombe mutants. The RAD2 gene increased survival of S. pombe rad13 to near the wild-type level after UV irradiation and had no effect on any of the other mutants tested. S. pombe rad13 mutants are somewhat defective in removal of pyrimidine dimers so complementation by the S. cerevisiae RAD2 gene suggests that the genes may code for equivalent proteins in the two yeasts.
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    URL: http://dx.doi.org/10.1007/BF00445748
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  • 85
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    Introduction of nonselectible 2μ plasmid into [cir°] cells of the yeast S. cerevisiae by DNA transformation and in vivo site-specific resolution (1989)
    Springer
    Current genetics 15 (1989), S. 83-90 
    Details
    ISSN: 1432-0983
    Keywords: DNA transformation ; Saccharomyces cerevisiae ; Site-specific recombination ; 2μ DNA plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The 2μ DNA plasmid of the yeast Saccharomyces cerevisiae does not confer any known selectable phenotype to the host cell carrying it. Selection of cells transformed with purified 2μ DNA therefore cannot be achieved, and the intracellular presence of 2μ can only be assessed by molecular analysis of the DNA complement. In addition, 2μ alone does not replicate in bacterial hosts, thus rendering its amplification by conventional methods impossible. We have isolated a shuttle plasmid, pBH-2L, generated by in vivo sites-pecific recombination between the endogenous 2μ DNA plasmid and pRL, a pBR322 derivative containing the yeast LEU2 gene and one 2μ repeat sequence associated with the origin of replication. This new shuttle plasmid has the property, when transformed into yeast, of undergoing site-specific recombinational resolution between its two direct repeat sequences. This releases 2μ plasmid and pRL as individual molecules. The latter can undergo progressive mitotic loss during growth in nonselective medium, ultimately leaving leucine auxotrophic transformants that contain only 2μ DNA plasmid. This system can be utilized to introduce 2μ DNA alone into cells lacking it, thereby providing a novel means to study the biology and the molecular genetics of the plasmid and its potential practical applications as a vector.
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    URL: http://dx.doi.org/10.1007/BF00435453
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  • 86
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    Heterologous gene expression of the glyphosate resistance marker and its application in yeast transformation (1989)
    Springer
    Current genetics 15 (1989), S. 91-98 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Vector ; Glyphosate resistance ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The E. coli aroA gene was inserted between yeast promoter and terminator sequences in different shuttle expression plasmids and found to confer enhanced EPSP synthase activity as well as resistance to glyphosate toxicity. Subsequently, a transformation system using these newly constructed vectors in yeast was characterized. The efficiency of the glyphosate resistance marker for transformation and selection with plasmid pHR6/20-1 in S. cerevisiae laboratory strain SHY2 was found to be relatively high when compared with selection for LEU2 prototrophy. The fate of the recombinant plasmid pHR6/20-1 in the transformants, the preservation of the aroA E. coli DNA fragment in yeast, mitotic stability, EPSP synthase activity, and growth on glyphosate-containing medium have been investigated. As this plasmid also allows direct selection for glyphosate resistant transformants on rich media, the glyphosate resistance marker was used for transforming both S. cerevisiae laboratory strain SHY2 and brewer's yeast strains S. cerevisiae var. “uvarum” BHS5 and BHS2. In all cases, the vector pHR6/20-1 was maintained as an autonomously replicating plasmid. The resistance marker is, therefore, suitable for transforming genetically unlabeled S. cerevisiae laboratory, wild, and industrial yeast strains.
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    URL: http://dx.doi.org/10.1007/BF00435454
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  • 87
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    A rapid and simple method for extracting yeast mitochondrial DNA (1989)
    Springer
    Current genetics 15 (1989), S. 235-237 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial DNA ; Method of extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid method for the extraction of yeast mitochondrial DNA (mtDNA) is described. In comparison with previous methods, it simplifies several steps, does not require either the isolation of mitochondria or phenol treatment and is less time consuming. This protocol gives a high yield of pure mtDNA (50–120 μg from a 100-ml culture), which can be directly used in various molecular applications: restriction enzyme digestion, electrophoresis, blotting, labeling, cloning and sequencing.
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    URL: http://dx.doi.org/10.1007/BF00435511
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  • 88
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    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.
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    URL: http://dx.doi.org/10.1007/BF00376739
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  • 89
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    A DNA sequence in Saccharomyces exiguus is homologous with the HO gene of Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 399-401 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces exiguus ; Saccharomyces cerevisiae ; HO gene ; MAT gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNA of Saccharomyces exiguus was analyzed by Southern hybridization using cloned MATa, MATα, and HO genes of Saccharomyces cerevisiae as probes. It was shown that S. exiguus has a DNA sequence homologous with the HO gene of S. cerevisiae and that this DNA sequence is on a chromosome of about 940 kb of DNA in S. exiguus. However, there is no DNA sequence in S. exiguus that is homologous with the MAT genes of S. cerevisiae.
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    URL: http://dx.doi.org/10.1007/BF00376794
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  • 90
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    Biosynthesis of the peroxisomal dihydroxyacetone synthase from Hansenula polymorpha in Saccharomyces cerevisiae induces growth but not proliferation of peroxisomes (1989)
    Springer
    Current genetics 16 (1989), S. 13-20 
    Details
    ISSN: 1432-0983
    Keywords: Peroxisomes ; Protein import ; Saccharomyces cerevisiae ; Hansenula polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DAS gene of Hansenula polymorpha was expressed in Saccharomyces cerevisiae under the control of different promoters. The heterologously synthesized dihydroxyacetone synthase (DHAS), a peroxisomal enzyme in H. polymorpha, shows enzymatic activity in baker's yeast. The enzyme was imported into the peroxisomes of S. cerevisiae not only under the appropriate physiological conditions for peroxisome proliferation (oleic acid media), but also in glucose-grown cells where it induced the enlargement of the few peroxisomes present. This growth process was not accompanied by an increase in the number of microbodies, which suggests a separate control mechanism for peroxisomal proliferation.
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    URL: http://dx.doi.org/10.1007/BF00411078
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  • 91
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    Molecular characterization of the two genes SNQ and SFA that confer Hyperresistance to 4-nitroquinoline-N-oxide and formaldehyde in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 65-74 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mutagen hyperresistance ; Southern, Northern analysis ; Gene transplacement ; Transposon mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genes SNQ and SFA confer hyperresistance to 4-NQO and FA when present on a multi-copy plasmid in yeast. Both are non-essential genes since transplacement of SNQ by a disrupted snq-0::LEU2 yielded stable and viable haploid integrants. Southern analysis revealed that SNQ and SFA are single-loci genes, and OFAGE analysis showed that they are located on chromosome XIII and IV, respectively. Northern blot analysis of SNQ and SFA revealed poly(A)+ RNA transcripts of 2 kb and 1.7 kb, respectively. Nuclease S 1 mapping showed SNQ to have a coding region of 1.6 kb and SFA, one of 1.3 kb. The 5′ coding regions were determined for both genes, while the 3′ end could only be determined for gene SNQ. Both genes do not appear to contain introns. The SFA locus was also mapped by transposon mutagenesis. Tn10-LUK integrants disrupted the SFA gene function at sites that were determined by subcloning to lie within the SFA transcription unit.
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    URL: http://dx.doi.org/10.1007/BF00393397
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  • 92
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    Reassessing the genotoxic potential of 8-MOP + UVA-induced DNA damage in the yeast Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 75-80 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Psoralen photoaddition ; Interstrand cross-link ; Repair deficiency ; Genotoxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two different UVA irradiation systems were initially biologically calibrated with two haploid yeast strains proficient and deficient, respectively, in nucleotide excision repair. The number of DNA lesions introduced into the cell's genome by the photoactivated bifunctional furocoumarin 8-MOP was then calculated by means of the applied UVA exposure doses. At LD37 the repair-proficient wild type had about 14 ICL and 34 furan-side monoadducts in its DNA, while doubly blocked repair mutant rad3-12 pso1-1 had 2 ICL and 3 monoadducts. Locus-specific reversion of lys1-1 followed two-hit kinetics in the repair-proficient wild type and one-hit kinetics in an excision-deficient rad2-20 mutant, as would be expected if ICL was the main type of mutagenic lesion in the wild type and monoadducts the main mutagenic lesion type in the excision-deficient strain. Quantitative comparison of 8-MOP + UVA-induced ICL with those induced by bifunctional mustard revealed the former to have a much higher genotoxicity.
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    URL: http://dx.doi.org/10.1007/BF00393398
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    Cloning and characterization of the SKI3 gene of Saccharomyces cerevisiae demonstrates allelism to SKI5 (1989)
    Springer
    Current genetics 16 (1989), S. 139-143 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; SKI3 ; SKI5 ; M1 dsRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified a mutant strain of the yeast Saccharomyces cerevisiae which overproduces killer toxin. This strain contains a single mutation which fails to complement defects in both the SKI3 and SKI5 genes, while a cloned copy of this gene complements both the ski3 and ski5 defects. The level of secreted toxin from a cDNA based plasmid is not increased in a ski3 strain, showing that the overproduction phenotype is dependent upon an increased level of M1 dsRNA.
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    URL: http://dx.doi.org/10.1007/BF00391469
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  • 94
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    High frequency recombination and the expression of genes cloned on chimeric yeast plasmids: Identification of a fragment of 2-μm circle essential for transformation (1980)
    Springer
    Current genetics 2 (1980), S. 17-251 
    Details
    ISSN: 1432-0983
    Keywords: Gene cloning ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have carried out experiments aimed at explaining the observed variations in transformation frequencies when Saccharomyces cerevisiae or Saccharomyces carlbergensis are transformed with chimeric plasmids that contain one of 4 possible EcoRI fragments of the yeast 2-μm circle. These plasmids fall into 2 classes when used to transform 2 different yeast his3 auxotrophs, one (strain LL20) harbours indigenous 2-μm circle, and the other (strain YF233) is devoid of this plasmid. Hybrid plasmids containing either the 2.4 mega-dalton (mD) R-form EcoRI fragment (pYF88) or the l.4 mD L-form EcoRI fragment (pYF177) of 2-μm circle transform either of the two hosts at a high frequency (50,000 colonies per Mg in LL20 and 10,000 colonies per μg in YF233). Hybrid plasmids containing the 1.5 mD R-form EcoRI fragment (pYF87) or the 2.5 mD L-form EcoRI fragment (pYF178) of the 2-μm circle transform LL20 at a reduced frequency (6,000–16,000 colonies per μg) and YF233 at extremely low frequencies (1–5 colonies per μg). All plasmids retrieved from strain YF233 that had been transformed with pYF88 or pYF177 were identical to the original transforming plasmid. Of the plasmids retrieved from strain LL20 that had been transformed with pYF87 and pYF178, approximately half had acquired an extra copy of the 2-μm circle. Of the plasmids retrieved from strain LL20 that had been transformed with pYF88 and pYF177, an average of only approximately 13% had acquired an extra copy of 2-μm circle. Taken together, these observations indicate that the transformation of yeast by a plasmid lacking the ability to replicate (pYF87 and pYF1780) occurs by the recombinational acquisition of 1 copy of the host 2-μm circle, which serves to supply the incoming plasmid with missing essential sequences. A comparison of 2-μm circle DNA fragments carried by pYF88 and pYF177 indicates that the region of 2-μm circle required for high frequency transformation is a 1.2 mD segment that is common to the 2.4 mD R-form and 1.4 ml) L-form EcoRI fragments. This region extends from the EcoRI cut site adjacent to the PstI site, through to the end of the inverted repeat. However, the inverted repeat sequence alone is not sufficient to bestow high frequency transformation of yeast.
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    URL: http://dx.doi.org/10.1007/BF00445690
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    Transcriptional units of GAL genes in Saccharomyces cerevisiae determined by ultraviolet light mapping (1980)
    Springer
    Current genetics 2 (1980), S. 223-228 
    Details
    ISSN: 1432-0983
    Keywords: Transcriptional Units ; GAL Genes ; Saccharomyces cerevisiae ; UV mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The size of the transcriptional unit of the structural genes for three galactose-metabolizing enzymes which form a cluster on chromosome II in Saccharomyces cerevisiae was studied by the ultraviolet light (UV)-mapping technique. Thus the size of the primary transcripts of GAL7 for galactose-1-phosphate uridylyl transferase, GAL10 for uridine diphosphoglucose 4-epimerase, or GAL1 for galactokinase were estimated to be 0.81 x 106, 1.1 x 106, or 1.3 x 106 respectively. In the light of these data together with the known directions of transcription of the genes, we concluded that each of three genes was transcribed from its own promoter.
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    Cloning of a DNA fragment from Cephalosporium acremonium which functions as an autonomous replication sequence in yeast (1984)
    Springer
    Current genetics 8 (1984), S. 155-163 
    Details
    ISSN: 1432-0983
    Keywords: Cephalosporium acremonium ; Mitochondrial DNA ; Autonomous replication sequence ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fragment of DNA which functions as an autonomous replication sequence in yeast was cloned from Cephalosporium acremonium. Mitochondrial DNA (mtDNA) was isolated from an industrial strain of C. acremonium (08G-250-21) highly developed for the production of the antibiotic, cephalosporin C. Size, 27 kb, and restriction pattern indicated this DNA was identical to mtDNA previously isolated (Minuth et al. 1982) from an ancestral strain (ATTC 14553) which produces very low amounts of cephalosporin C. A 1.9 kb Pst1 fragment of the Cephalosporium mtDNA was inserted into a Pst1 site of the yeast integrative plasmid, Ylp5, to produce a 7.5 kb plasmid, designated pPS1. The structure of pPS1 was verified by restriction analysis and hybridization. PS1 transformed Saccharomyces cerevisiae (DBY-746) to uracil prototrophy at a frequency of 272 transformants/μg DNA. Transformation frequencies of 715 transformants/μg DNA and zero were obtained for the replicative plasmid, YRp7, and the integrative plasmid YIp5, respectively. Southern hybridization and transformation of E. coli by DNA from yeast transformed by pPS1 verified that pPS1 replicates autonomously in yeast. The uracil-independent pPS1-yeast transformants were mitotically unstable. The average retention of pPS1 after three days growth in selective and non-selective medium was 4.5% and 0.4%, respectively, compared to retentions of 4.6% and 0.5% for YRp7. The properties of pPS1 were compared to those of a related plasmid, pCP2. pCP2 was constructed (Tudzynski et al. 1982) by inserting the C. acremonium 1.9 kb Pst1 fragment into the yeast integrative plasmid, pDAM1.
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    URL: http://dx.doi.org/10.1007/BF00417811
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    Structure and function of the TRP3 gene of Saccharomyces cerevisiae: Analysis of 3′- and 5′-deletions in vivo and in vitro (1984)
    Springer
    Current genetics 8 (1984), S. 173-180 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; TRP3 gene ; Deletion analysis ; Enzyme function
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two sets of deletions, entering the TRP3 gene of Saccharomyces cerevisiae from the 3′- and the 5′-end were constructed. Complementation analysis with chromosomal trp3A, trp3B and trp3C mutations was done by introducing the 3′- and 5′-truncated gene on a multicopy 2 μm-vector. The N-terminal glutamine amido transferase function is encoded by a DNA fragment of 600–700 bp, and the C-terminal indole-3-glycerol-phosphate synthase function by a DNA fragment of about 900 bp, whereas both functions together are encoded by a contiguous DNA fragment of about 1,500 bp. The bi functional TRP3-peptide thus could be dissected into two catalytically independent peptides in vivo. For the indole-3-glycerol-phosphate synthase activity, independent catalytic activity was also demonstrated in vitro: deletions entering the TRP3 gene from the 5′-end, and lacking large parts of the sequence coding for the glutamine amidotransferase function, still are able to ex press a peptide exhibiting functional indole-3-glycerol phosphate synthase activity in vitro. Deletion plasmids pME505·De1C102·2μm and DelC10·2μm exhibited shorter TRP3 transcripts according to the deleted DNA-fragments (150 and 426 by respectively) but yielded peptides of invariable Mr of 35,000 d. Transcription and translation of these peptides, which probably represent the independently folding indole-3-glycerol-phosphate synthase core are discussed.
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    URL: http://dx.doi.org/10.1007/BF00417813
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    Interactions among genes controlling sensitivity to radiation (RAD) and to alkylation by nitrogen mustard (SNM) in yeast (1982)
    Springer
    Current genetics 5 (1982), S. 33-38 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Multiple mutants of DNA repair ; Sensitivity to nitrogen mustard and to radiation ; Thermoconditional DNA repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three haploid yeast mutants (snm) sensitive or thermoconditionally sensitive to the DNA cross-linking agent nitrogen mustard (HN2) were crossed with four rad strains representing mutations in the three pathways of DNA dark repair. The resulting haploid double and triple mutant strains were tested for their sensitivity to UV, HN2 and HN1. From the observed epistatic or synergistic interactions of the combinations of mutant alleles we could derive the relation of the SNM1 and SNM2 genes to the postulated repair pathways. Alleles snm1-1 and snml-2 ts were found epistatic to genes of the rad3 group, whereas snm2-1 ts was epistatic to rad6. The snm1 and snm2 mutant alleles interacted synergistically. From these data it is concluded that the SNM1 gene product plays a cross-link specific role in excision repair while the SNM2 gene product may be involved in a system of error-prone repair.
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    URL: http://dx.doi.org/10.1007/BF00445738
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    Isolation of the TRP2 and the TRP3 genes of Saccharomyces cerevisiae by functional complementation in yeast (1982)
    Springer
    Current genetics 5 (1982), S. 39-46 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; TRP2 gene ; TRP3 gene ; Cloning in yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper describes the isolation of the TRP2 and the TRP3 genes of Saccharomyces cerevisiae. Two pools of plasmids consisting of BamHI and Sa1GI yeast DNA inserts into the bifunctional yeast — Escherichia coli vector pLC544 (Kingsman et al. 1979) were constructed in E. coli and used for the isolation of the two genes by selection for functional complementation of trp2 and trp3 mutations, respectively, in yeast. The TRP2 gene was isolated on a 6.2 kb BamHl and a 5.8 kb Sa1GI yeast DNA fragment which shared an identical 4.5 kb BamHI-SaIGI fragment. The TRP3 gene was located on a 5.2 kb BamHl fragment. By physical, genetic and physiological experiments it could be shown that the cloned yeast DNA fragments contained the whole structural sequences as well as the regulatory regions of the TRP2 and the TRP3 genes.
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    URL: http://dx.doi.org/10.1007/BF00445739
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  • 100
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    Purification and genetic control of α-pheromone-inactivating glycoproteins in the yeast Saccharomyces cerevisiae (1984)
    Springer
    Current genetics 9 (1984), S. 39-45 
    Details
    ISSN: 1432-0983
    Keywords: α-Pheromone-inactivating glycoproteins ; bar1-1 ; Barrier proteins ; Purification ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two kinds of a-mating-type-specific proteins inactivating α pheromone (α factor) were purified from heat shock extract of MATa cells. Their molecular weights were estimated to be 400,000 and 200,000 by gel filtration. Both proteins were detected in MATa SST1 cells but not in MATα SST1, MATa sst1-1 and MATa/MATα SST1/SST1 cells. In addition, the proteins were detected in matα2-1 SST1 cells but not in matα1-2 SST1 cells. From these results, it is concluded that these proteins are synthesized under the control of the SST1 gene and responsible for the Barrier action of MATa cells. The relationship of these proteins to the secreted Barrier protein having a higher molecular weight is discussed.
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    URL: http://dx.doi.org/10.1007/BF00396202
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