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  • Saccharomyces cerevisiae
  • stability
  • Springer  (142)
  • American Association for the Advancement of Science
  • American Institute of Physics (AIP)
  • 1980-1984  (96)
  • 1975-1979  (46)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Computing 24 (1980), S. 341-347 
    ISSN: 1436-5057
    Keywords: Numerical analysis ; Volterra integral equations of the second kind ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science
    Description / Table of Contents: Zusammenfassung Ziel dieser Arbeit ist es, die Stabilitätseigenschaften einer Klasse Volterrascher Integralgleichungen zweiter Art zu untersuchen. Unsere Behandlung ist der üblichen Stabilitätsanalyse ähnlich, in der die Kernfunktionen zu einer im voraus beschränkten Klasse von Testfunktionen gehören. Wir haben die Klasse der “endlich zerlegbaren” Kerne betrachtet. Stabilitätsbedingungen werden abgeleitet und verglichen mit den Bedingungen für die einfache Testgleichung. Es zeigt sich, daß die neuen Kriteria einschränkender sind als die konventionellen Bedingungen. Der praktische Wert wird getestet durch numerische Experimente mit der Trapezregel.
    Notes: Abstract The purpose of this paper is to analyse the stability properties of a class of multistep methods for second kind Volterra integral equations. Our approach follows the usual analysis in which the kernel function is a priori restricted to a special class of test functions. We consider the class of finitely decomposable kernels. Stability conditions will be derived and compared with those obtained with the simple test equation. It turns out that the new criteria are more severe than the conventional conditions. The practical value is tested by numerical experiments with the trapezoidal rule.
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  • 2
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    Springer
    European journal of nutrition 22 (1983), S. 205-212 
    ISSN: 1436-6215
    Keywords: Schwermetallwirkung ; Malatdehydrogenase ; Glutamatdehydrogenase ; Glycerinaldehyd-3-phosphatdehydrogenase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The difference between cadmium, zinc, lead, and mercury in regard of their effects on the activity of the enzymes tested is very slight. Concentrations higher than 10−5 M reduce significantly the activity of the enzymes, and concentrations of approximately 10−3 M inhibit it completely. An increase of the activity cannot be detected. The addition of combinations of cadmium, zinc, and lead results in a summing up of the toxic effects, whereas the interaction between mercury and the other three heavy metals shows a cumulative effect, which is appointed nearly completely by the heavy metal more toxic. The findings suggest that under in-vitro conditions there exists a direct interaction between the heavy metals and the enzymes.
    Notes: Zusammenfassung Die vier Schwermetalle Cadmium, Zink, Blei und Quecksilber unterscheiden sich in ihrer Wirkung auf die Aktivität der untersuchten Enzyme nur sehr wenig. Konzentrationen über 10−5 M vermindern die Enzymaktivität signifikant, und Konzentrationen von etwa 10−3 M unterbinden sie völlig. Eine Steigerung der Enzymaktivität läßt sich nicht feststellen. Die Zugabe von Cadmium-, Zink- und Bleikombinationen führt zu einer Addition der toxischen Effekte, während bei der Interaktion zwischen Quecksilber und den anderen drei Schwermetallen die Gesamtwirkung fast ausschließlich durch das stärker hemmende Schwermetall allein bestimmt wird. Die erhaltenen Ergebnisse lassen vermuten, daß es unter Invitro-Bedingungen zu einer direkten Wechselwirkung zwischen den Schwermetallen und den Enzymen kommt.
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  • 3
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    Springer
    Rheologica acta 22 (1983), S. 284-290 
    ISSN: 1435-1528
    Keywords: Viscometric flow ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Physics
    Notes: Abstract This paper examines three-dimensional disturbances of a plane steady shear flow of simple fluids with short memory. Under the assumption of nearly-viscometric flow, constitutive equations are derived and then a general form of the Reynolds-Orr energy equation is obtained. With the aid of this derived energy formula, sufficient conditions are generated for the stability of three-dimensional disturbances of the planar viscometric flow. These conditions are analysed and a comparison is made with the corresponding two-dimensional stability problem. There is a strong indication that the basic flow is less stable against three-dimensional disturbances than against two-dimensional ones.
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  • 4
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    Springer
    BIT 17 (1977), S. 321-328 
    ISSN: 1572-9125
    Keywords: 5.15 ; nonlinear equation ; root finding ; multiple root ; secant method ; Steffensen procedure ; order of convergence ; efficiency ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics
    Notes: Abstract A superlinear procedure for finding a multiple root is presented. In it the secant method is applied to the given function divided by a divided difference whose increment shrinks toward zero as the root is approached. Two function evaluations per step are required, but no derivatives need be calculated.
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  • 5
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    Springer
    Cellular and molecular life sciences 40 (1984), S. 1159-1161 
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; 5-trifluoromethyl-6-àzauracil ; yeast cell cultures ; cell division ; inhibition of
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cell division, as studied in asynchronous cultures of yeast cells, is sensitive to 5-trifluoromethyl-6-azauracil (F3CAzU). Under defined conditions (10 mmoles l−1 F3CAzU) this compound blocks immediately and completely the process of cell division. Using synchronized cells, the time-point at which division process of yeast cell can be inhibited by F3CAzU has been determined. The inhibitor effect of this compound is completely reversed by thymine, thymidine and uracil.
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  • 6
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    Archive for rational mechanics and analysis 72 (1980), S. 203-218 
    ISSN: 1432-0673
    Keywords: simple fluid ; viscoelastic ; fading memory ; stability ; Liapunov function ; dynamical system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics , Physics
    Notes: Abstract The nonlinear equations of motion for an incompressible simple fluid, occupying a fixed bounded container, are formulated on the basis of the “finitelinear” viscoelasticity theory for materials with fading memory; this formal boundary-initial value problem is then viewed as a nonlinear abstract evolution equation on a certain Hilbert space. It is shown that a linearized version of this evolution equation is associated with a linear dynamical system on this Hilbert space, and several results for stability and asymptotic behavior for this linearized problem are proved through the use of Liapunov stability methods. On the assumption that the original nonlinear evolution equation also is associated with some dynamical system on the same space, it is shown that the rest condition of the fluid is stable and all motions are bounded. The Liapunov function employed for this purpose can be interpreted as a mechanical energy function for the fluid.
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  • 7
    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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  • 8
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    Springer
    Current genetics 1 (1979), S. 63-74 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Translation ; Coordinate regulation ; Electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The products of protein synthesis from exponential phase cultures of Saccharomyces cerevisiae grown at 23 °C or at 36 °C appear to be essentially identical. However, yeast cells respond to a shift in culture temperature from 23 °C to 36 °C with the rapid de novo synthesis of a polypeptide species of molecular weight 100,000. Within 60–90 min after the shift this polypeptide represents approximately 2.5% of the total cellular protein, a 5–10 fold increase over the preshift level. The level of this polypeptide then decreases with continued growth of the cells at 36 °C. Analyses by SDS-polyacrylamide gel electrophoresis of polypeptides obtained from cells pulse labeled with [35S]methionine demonstrate that following a temperature shift from 23 °C to 36 °C the synthetic rate of the 100,000 molecular weight polypeptide (as well as a number of other polypeptide species) increases to a level at least 10 fold higher than that observed prior to the shift. A concomittant decrease is observed in the synthesis of a large number of polypeptide species which were actively synthesized before the shift. Maximum changes in synthetic rates are observed 20–30 min after the shift and preshift synthetic patterns are regained within 60–90 min. Synthetic changes of the same magnitude and time course can be produced by short (20–30 min) exposures to 36 °C implicating a heat shock response. Several of the transiently induced polypeptides, including the 100,000 molecular weight species, show an affinity for DNA as determined by DNA-cellulose chromatography.
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  • 9
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    Current genetics 2 (1980), S. 115-120 
    ISSN: 1432-0983
    Keywords: Galactose fermentation ; Saccharomyces cerevisiae ; Regulatory mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A novel type of regulatory mutation for galactose metabolism in Saccharomyces cerevisiae is described. The mutation named gal11 was recessive, non-allelic to GAL4, GAL80, GAL2, or GAL3, and unlinked to the gene cluster of GAL1, GAL10, and GAL7. It caused a ‘coordinate’ reduction of galactokinase, galactose-1-P uridylyl transferase, and UDP-glucose 4-epimerase by a factor of more than 5, rendering the mutant cells galactose-nonfermenting. The effect of the mutation was manifested not only in cells grown on galactose but also in cells constitutively synthesizing the galactose-metabolizing enzymes.
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  • 10
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    Current genetics 2 (1980), S. 223-228 
    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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  • 11
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    Computing 31 (1983), S. 261-267 
    ISSN: 1436-5057
    Keywords: 65M10 ; Dispersive equation ; finite difference ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science
    Description / Table of Contents: Zusammenfassung Dieser Artikel beinhaltet eine Zusammenstellung von Differenzenverfahren für die Dispersionsgleichungu 1=au xxx. Es werden Kriterien zur Herleitung von Stabilitätsbedingungen für Differenzenverfahren angegeben und auf die angegebenen Differenzenverfahren angewendet.
    Notes: Abstract In this paper a table of difference schemes for the dispersive equationu i=au xxx is presented. A collection of criterions for deriving stability conditions of difference schemes is given and applied to these difference schemes.
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  • 12
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    Computing 32 (1984), S. 229-237 
    ISSN: 1436-5057
    Keywords: 65L05 ; 65L07 ; Stiff system ; Rosenbroek method ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science
    Description / Table of Contents: Zusammenfassung In dieser Arbeit wird die Stabilität des Kaps-Rentrop-Verfahrens in die Anwesenheit nichtlinearer Steifheit (Stiffness) analysiert. Dazu werden mittels eines einfachen Modells zwei Größen introduziert. Die Werte dieser Größen reflektieren gewissermaßen das Verhalten eines Kaps-Rentrop-Verfahrens in die Anwesenheit einer bestimmten Kopplung zwischen die beiden Komponenten in das steife System gewöhnlicher Differentialgleichungen. Einige numerische Beispiele veranschaulichen die Analyse.
    Notes: Abstract In this paper we give an analysis of the effect of stiff nonlinearities on the behavior of a Kaps-Rentrop method. To that end we introduce two quantities related to a simple model. The values of these quantities determine to some extent the behavior of a Kaps-Rentrop method in case of a strong coupling between the smooth component and the transient one. Numerical examples illustrate the theoretical results.
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  • 13
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    Journal of statistical physics 14 (1976), S. 399-416 
    ISSN: 1572-9613
    Keywords: Reservoir-driven open systems ; coherent states ; entropy production ; nonlinear equations ; irreversible processes ; stationary state ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract A kinetic equation for the density matrix of a monomode laser with explicit coupling with a thermal reservoir representing the cavity and a nonthermal one representing the pumping mechanism is derived. The macroscopic behavior of this system, inferred from Glauber's P function, is discussed within the framework of Glansdorff-Prigogine's theory of far-from-thermal-equilibrium open systems.
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  • 14
    ISSN: 1572-9613
    Keywords: Correlation inequalities ; classical and quantum continuous systems ; positive type potentials ; stability ; thermodynamic limit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract We study quantum mechanical systems of particles with Bose or Fermi statistics interacting via two-body potentials of positive type in thermal equilibrium. We rewrite partition functions, reduced density matrices (RDMs), and correlation functions in terms of Wiener and Gaussian functional integrals (sine-Gordon transformation). This permits us, e.g., to apply correlation inequalities. Our main results include an analysis of stability versus instability in the grand canonical ensemble and, for charge-conjugation-invariant systems, upper and lower bounds on RDMs, the existence of the thermodynamic limit of pressure, RDMs and correlation functions, an inequality comparing correlations with Fermi statistics to ones with Bose statistics, and inequalities which are important in the study of Bose-Einstein condensation and of superconductivity.
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  • 15
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    Applied microbiology and biotechnology 16 (1982), S. 75-80 
    ISSN: 1432-0614
    Keywords: Immobilization of yeast ; Saccharomyces cerevisiae ; Ethanol production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Baker's yeast (Saccharomyces cerevisiae) was immobilized in gels made of prepolymerized, linear, water soluble polyacrylamide, partially substituted with acylhydrazide groups. Gelation was effected by the addition of controlled amounts of dialdehydes (e.g. glyoxal). The immobilized yeasts retained full glycolytic activity. Moreover, the entrapped cells were able to grow inside the chemically corsslinked gel during continuous alcohol production. Glyoxal was found to be the most favourable crosslinking agent for this system. the system employed allowed for the free exchange of substrate and products. The gel surrounding the entrapped cells had no effect on temperature stability profile. On the other hand, substantial enhancement in survival of cells in presence of high ethanol concentrations was recorded for the entrapped yeast. The capability of the immobilized yeast to carry out continuous conversion of glucose to ethanol was demonstrated.
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  • 16
    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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  • 17
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    Current genetics 7 (1983), S. 165-166 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cephalosporium acremonium ; Mitochondrial hybrid vector ; Nuclear association
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hybrid vector pCP2, consisting of the bacterial plasmid pBR325, the nuclear gene Leu-2 of Saccharomyces cerevisiae and a fragment of mitochondrial DNA from Cephalosporium acremonium, was found to associate with the nucleus in a transformed strain of Saccharomyces cerevisiae. This was inducted by (1) efficient expression of the Leu-2 gene as evidenced by a short generation time on selective medium; (2) independence of Leu-2 gene expression from mitochondrial protein synthesis, since pCP2 was shown to replicate and to be expressed in petite mutants; (3) association of pCP2 with isolated DNA from nuclei as proved by transformation experiments with E. coli.
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  • 18
    ISSN: 1432-0983
    Keywords: Catalase ; Saccharomyces cerevisiae ; Heme ; Posttranscriptional control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Compared to wild type cells, strains bearing the pleiotropic regulatory mutations cgr4 or cas1 synthesize apocatalase T at a high rate when grown on high glucose. Like heme-deficient ole3 single mutants, ole3 cgr4 and ole3 cas1 double mutants accumulate no catalase T protein in vivo. This defect introduced by the ole3 mutation is cured by the addition of ALA. By use of the inhibitor actinomycin D we confirm previous findings that ole3 mutants lack catalase T mRNA and show that (i) the ole3 cgr4 and ole3 cas1 double mutants do accumulate catalase T mRNA or mRNA precursor, and (ii) the processing or translation of this RNA or the accumulation of apocatalase T depends on the presence of home.
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  • 19
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; G1 cdc mutants ; tα-factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants in four G1 cdc strains of Saccharomyces cerevisiae were isolated which failed to show division arrest in the presence of α-factor. The cell cycle properties, terminal arrest morphology and mating competence of these mutants at the restrictive temperature were examined. The G1 specific arrest of the cdc 36 and cdc39 mutants is dependent upon the availability of an intact mating factor response system in Mat a cells. Cdc28 and cdc37 mutants exert their cell cycle blocks independently of the mating factor pathway. It is likely that the nature of the primary growth defect in cdc36 and cdc39 mutants is such that the α-factor pathway is activated in the absence of the pheromone at the restrictive temperature and that G1 arrest is a secondary consequence of a non-cycle specific event in such mutants.
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  • 20
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    Current genetics 7 (1983), S. 235-237 
    ISSN: 1432-0983
    Keywords: DNA replication ; Shuttle vectors ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitotic segregation of three 2 μm-pBR322 chimaeric plasmids (YEp6, YEp21, and YEp24) was studied in yeast. Each displayed a characteristic rate of loss: YEp6 was lost at approximately twice the rate of YEp21 and YEp24. The loss rates were not significantly increased when two chimaeric plasmids were coresident, nor was the endogenous 2 μm plasmid itself displaced. Therefore these plasmids appear to be compatible in yeast.
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  • 21
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    Current genetics 8 (1984), S. 559-566 
    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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  • 22
    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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  • 23
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    Current genetics 4 (1981), S. 85-90 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Nuclear isolation ; Percoll ; in vitro Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A procedure has been developed for the rapid isolation of yeast nuclei in high yield using Percoll gradients. The nuclei are substantially free of cytoplasmic contamination as measured by alcohol dehydrogenase activities, have the typical chromatin digestion pattern when digested with nucleases, are useful for isolation of nuclear proteins and for in vitro transcription experiments.
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  • 24
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; DEL1 ; rad ; ste7
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In DEL1 strains of the yeast, Saccharomyces cerevisiae, the iso-1-cytochrome c (CYC1) region is flanked on either side by Tyl elements in direct orientation which promote cyc1 deletions of the bracketed DNA in the haploid cell. In this study, we asked which genes might control this event by testing the possibility that the DEL1 mutation mechanism requires an enzyme (or enzymes) that is also utilized in the repair of damaged DNA. To this end, we independently coupled eight repair mutations, rad3–2, rad4–4, rad6–1, rad6–3, rad9–1, rev3–1, rad50–1, and rad51-1, toDEL1 and asked whether DEL1 was still functional. We found that none of these rad mutations significantly affects the mutation frequency of 10−6-10−5 established in DEL1 strains for the CYC1 locus. Furthermore, we determined that ste7, a temperature-sensitive sterile allele known to alter gene regulation in Ty-mediated mutations, is not required for DEL1 function. Finally, DEL1 is not temperature-sensitive at 23° or 37 °C.
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  • 25
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    Current genetics 7 (1983), S. 369-377 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Isoenzymes ; Induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Tetrad analysis indicates that α-isopropylmalate synthase activity of yeast is determined by two separate genes, designated LEU4 and LEU5. LEU4 is identified as a structural gene. LEU5 either encodes another α-isopropylmalate synthase activity by itself or provides some function needed for the expression of a second structural gene. The properties of mutants affecting the biosynthesis of leucine and its regulation suggest that the expression of LEU1 and LEU2 (structural genes encoding isopropylmalate isomerase and β-isopropylmalate dehydrogenase, respectively) is controlled by a complex of a-isopropylmalate and a regulatory element (the LEU3 gene product). Similarities and differences between yeast and Neurospora crassa with respect to leucine biosynthesis are discussed.
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  • 26
    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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  • 27
    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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  • 28
    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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  • 29
    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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  • 30
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    Current genetics 6 (1982), S. 163-165 
    ISSN: 1432-0983
    Keywords: Hybridization ; Polyethylene glycol ; Nuclear transfer ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Summary Viable hybrids of Saccharomyces cerevisiae were obtained by transfer of isolated diploid nuclei into haploid protoplasts using a polyethylene glycol (PEG) fusion procedure.
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  • 31
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    Current genetics 7 (1983), S. 393-397 
    ISSN: 1432-0983
    Keywords: Trehalose ; Glycogen ; Sporulation ; Germination ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants with specific lesions were used to differentiate between the functions of glycogen and trehalose in S. cerevisiae. Diploids which harbor the glc1/glc1 mutation depend upon the phosphorylated, less active form of glycogen synthase and show a more active, phosphorylated form, of the enzyme trehalase. These conditions are due to a lesion in the regulating subunit of the cAMP-dependent protein kinase. Such cells are unable to sporulate. Diploids which contain the sst1/sst1 mutation have normal glycogen metabolism but their trehalose-6-phosphate synthase is not active. Such strains sporulate but germination is poor and only one-spore tetrads are formed. These results confirm that glycogen is needed to trigger sporulation while trehalose plays a role in the germination process. Different systems, I and II, of trehalose accumulation were proposed. System I would require the UDPG-linked trehalose synthase, whereas system II would constitute an alternative pathway, specifically induced or activated by the expression of a MAL gene. The presence of system II in its constitutive form in the constructed diploids would favour trehalose synthesis during growth on glucose, however, it did not overcome the glycogen deficiency during sporulation nor the lack of trehalose for germination. It seems that only system I, namely trehalose 6-P-synthase, plays a role in the germination process.
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  • 32
    ISSN: 1432-0983
    Keywords: Oversecretion mutants ; Protease defect ; Wall glucan defect ; Saccharomyces cerevisiae
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    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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  • 33
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; 5-aminolevulinate synthase ; Cloned gene
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    Topics: Biology
    Notes: Summary We have cloned the structural gene HEM1 for 5-aminolevulinate (ALA) synthase from Saccharomyces cerevisiae by transformation and complementation of a yeast hem1–5 mutant which was previously shown to lack ALA synthase activity (Urban-Grimal and Labbe Bois 1981) and had no immunodetectable ALA synthase protein when tested with yeast ALA synthase antiserum. The gene was selected from a recombinant cosmid pool which contained wild-type yeast genomic DNA fragments of an average size of 40 kb. The cloned gene was identified by the restauration.of growth on a non fermentable carbon source without addition of exogenous ALA. Sub cloning of partial Sau3A digests and functional analysis by transformation allowed us to isolate three independent plasmids, each carrying a 6 kb yeast DNA fragment inserted in either orientation into the single BamHI site of the vector pHCG3 and able to complement hem1–5 mutation. Analysis of the three plasmids by restriction endonucleases showed that HEM1 is contained within a 2.9 kb fragment. The three corresponding yeast trans formants present a 1, 2.5 and 16 fold increase in ALA synthase activity as compared to the wild-type strain. The gene product immunodetected in the transformant yeast cells has identical size as the wild-type yeast ALA synthase and its amount correlates well with the increase in ALA synthase activity.
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  • 34
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    Current genetics 6 (1982), S. 159-162 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Transformation ; Gene subcloning
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    Notes: Summary We have developped a procedure to delimit the boundaries of a cloned gene carried on a DNA fragment as large as 4 to 5 kilobases. The method consists in the following. Two series of limit digest products generated with a tetranucleotide recognition sequence endonuclease and originating from either of the two ends of this DNA segment are tested for their complementing capacity by yeast transformation. The gene is then delimited by the overlap of the two shortest complementing fragments.
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  • 35
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    Current genetics 9 (1984), S. 107-111 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; 2 μm minichromosomes ; Metrizamide gradients
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    Topics: Biology
    Notes: Summary Two micrometer minichromosomes from Saccharomyces cerevisiae were isolated without detergent using metrizamide gradients. 2 μm minichromosomes showed a lower density in metrizamide gradients relative to genomic chromatin. Our results suggest a lower ratio of proteins to DNA in 2-μm minichromosomes as compared with genomic chromatin. The procedure described herein yields minichromosomes free of cellular chromatin and ribosomal protein contamination. This method may be useful for the isolation and characterization of other yeast minichromosomes.
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  • 36
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    Current genetics 7 (1983), S. 427-431 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; DNA ; Alkaline elution
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    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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  • 37
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    Current genetics 8 (1984), S. 81-84 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial genes ; Vegetative segregation ; Uniparental inheritance
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    Topics: Biology
    Notes: Summary Zygotes of Saccharomyces cerevisiae that are heteroplasmic for mitochondrial alleles produce diploid progeny that are homoplasmic for one allele or the other, judged by the criterion that upon further subcloning they produce daughter cells of only one phenotype or the other. Here we show that when such cells are subjected to strong selection for the missing allele, it cannot be detected, so that it is probably not present in even a single copy.
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  • 38
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    Current genetics 4 (1981), S. 135-143 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondria ; Gene cloning ; Transfer RNA
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    Topics: Biology
    Notes: Summary We have studied the organization and expression of a group of tRNA genes located on a 2,700 base pair portion of the yeast mitochondrial genome between the genetic markers cap (chloramphenicol resistance) and oxil (cytochrome oxidase subunit II). This region is spanned by mitochondrial DNA inserts of two recombinant plasmids, pYm162 and pYm267, which have been extensively mapped and sequenced. This tRNA group is composed of six tRNA genes, coding for tRNA AAR Lys , tRNA AGR Arg , tRNA GGN Gly , tRNA GAY Asp , tRNA AGY Ser , and tRNA CGN Arg . We report the sequence for the majority of the 2,700 base pair region including the genes for all six tRNAs. All six genes are oriented in the same direction and are, therefore, transcribed from the same DNA strand. Further, a comparison of the organization of this region with the analogous region of a related wild type strain shows that the tRNA gene order in the two strains is the same. Five of the six tRNA genes have corresponding transcripts in wild type RNA. Although a potential structural gene for tRNA CGN Arg is present, we do not detect a tRNA CGN Arg gene transcript.
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  • 39
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; TRP2 gene ; TRP3 gene ; Cloning in yeast
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    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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  • 40
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    Current genetics 7 (1983), S. 433-438 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Yeast transformation ; Yeast autonomously replicating sequences ; Ribosomal RNA genes
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    Topics: Biology
    Notes: Summary We have previously demonstrated that the loss of Rcp-CEN3, a centromeric plasmid containing yeast rDNA autonomously replicating sequences (ARS) is as high as around 50% per generation for most yeast strains. In this study we have attempted to elucidate mechanisms underlying the high mitotic instability of Rcp-CEN3. For this purpose a tandem duplication of a rDNA ARS was constructed in Rcp-CEN3. The new plasmid having two ARSs possesses a markedly higher mitotic stability as compared to a monoARS Rcp-CEN3. The mitotic stability of this centromere-containing plasmid which has two replicators corresponds to the calculated value for the mitotic stability of two monoARS plasmids Rcp-CEN3 in given cells. Genetic analysis has demonstrated that both plasmids having one or two ARSs are maintained in the single copy state. These results demonstrate that the mitotic instability of centromeric plasmid Rcp-CEN3 carrying a rDNA ARS is associated with the absence of stringent control of replication from the rDNA ARS. A possible mechanism of replication of the chromosomal rDNA repeats in yeast is discussed in the light of this data.
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  • 41
    ISSN: 1432-0983
    Keywords: Iso-1-cytochrome c ; Saccharomyces cerevisiae ; Heme ; Transcription
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    Notes: Summary A Saccharomyces cerevisiae mutant (hem1 cycl-1) was transformed with plasmids bearing a chromosomal centromer (CEN3) and a 2 μm DNA replication origin. In one of the plasmids a functional CYC1 gene was present, in a second plasmid an XhoI fragment located between bases -245 and -678 upstream from the translation initiation codon had been deleted, in a third plasmid this region had been inverted. Results of hybridization experiments carried out with mRNA isolated from heme-deficient and heme-containing transformants indicated that heme controls transcription of the CYC1 gene and that DNA sequences located within the upstream XhoI fragment are involved in activation of the gene by heme.
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  • 42
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; TRP3 gene ; Sequence analysis ; Enzyme function
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    Notes: Summary The structure and function of the TRP3 gene of Saccharomyces cerevisiae were analyzed. Subcloning of an original 4.8 kb BamHI DNA fragment, carrying the yeast TRP3 gene, allowed for a localization of the gene on a 2.5 kb ClaI/BamHI fragment. Transcription was found to proceed from the ClaI site towards the BamHI site. Three major transcription start sites were determined at positions −92, −87, and −81 by S1-mapping. The synthesis of the TRP3 gene is regulated by the general control, and was found to take place- at the transcriptional level. The sequence of the 5′-noncoding region up to position −400 and part of the coding region to position 840 were determined. The 5′-noncoding region contains sequences common to most amino acid biosynthetic genes known so far, namely a presumptive ribosome binding site, “Goldberg-Hogness boxes”, and a consensus sequence, possibly involved in the general control. For the coding region a single open reading frame was found. The deduced amino acid sequence was aligned with homologous amino acid sequences of Neurospora crassa, Pseudomonas putida and Escherichia coli. The exceptionally high homology (40–60%) between these sequences led us to postulate that the TRP3 gene product is of the structure NH2-glutamine amidotransferase-indole-3-glycerol-phosphate synthase-COOH.
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  • 43
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cloning ; Suppressor
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    Notes: Summary A plasmid, pYsup1-1, containing a DNA fragment able to suppress the recessive mutant phenotype of the suppressor locus sup1 (allele sup1-ts36) of Saccharomyces cerevisiae was isolated from a bank of yeast chromosomal DNA cloned in cosmid p3030. The complementing gene was localized on a 2.6 kb DNA fragment by further subcloning. Evidence is presented that the cloned DNA segment codes for the sup1 structural gene (chromosome IIR).
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  • 44
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    Current genetics 8 (1984), S. 575-580 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Candida utilis ; Protoplast fusion
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    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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  • 45
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    Archives of microbiology 107 (1976), S. 207-214 
    ISSN: 1432-072X
    Keywords: Anthranilate synthase ; Cell permeabilisation ; Indoleglycerolphosphate synthase ; Saccharomyces cerevisiae ; Tryptophan biosynthetic enzymes ; Tryptophan pool
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    Notes: Abstract The free tryptophan pool and the levels of two enzymes of tryptophan biosynthesis (anthranilate synthase and indoleglycerolphosphate synthase) have been determined in a wild type strain of Saccharomyces cerevisiae and in mutants with altered regulatory properties. The tryptophan pool of wild type cells growing in minimal medium is 0.07 μmole per g dry weight. Addition of anthranilate, indole or tryptophan to the medium produces a fifteen- to forty-fold increase in tryptophan pool, but causes no repression of the biosynthetic enzymes. Inclusion of 5-methyltryptophan in the growth medium causes a reduction in growth rate and a derepression of the biosynthetic enzymes, and this is shown here not to be correlated with a decrease in the free tryptophan pool. Mutants with an altered anthranilate synthase showing decreased sensitivity to inhibition by l-tryptophan or by the analogue dl-5-methyltryptophan have a tryptophan pool far higher than the wild type strain, but no repression of indoleglycerolphosphate synthase was observed. Mutants with an anthranilate synthase more sensitive to tryptophan inhibition show a slightly reduced tryptophan pool, but no derepression of indoleglycerolphosphate synthase was found. A mutant with constitutively derepressed levels of the biosynthetic enzymes shows a considerably increased tryptophan pool. Addition of 5-methyltryptophan to the growth medium of non-derepressible mutants causes a decrease in growth rate accompanied by a decrease in the tryptophan pool.
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  • 46
    ISSN: 1432-072X
    Keywords: Yeasts ; Sugars ; d-Glucose ; 2-Deoxy-d-glucose ; Pichia pinus ; Transport ; Starvation ; Exponential growth ; Methodology ; Candida utilis ; Saccharomyces cerevisiae ; Rhodosporidium toruloides
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Some methods for measuring the uptake of sugars by yeasts were investigated critically. A study was made of the effects of starvation of Pichia pinus, Candida utilis, Saccharomyces cerevisiae and Rhodosporidium toruloides on their uptake of d-glucose and 2-deoxy-d-glucose. Marked changes in the rates of uptake of these sugars occurred during 10 h of starvation, including (a) an immediate increase of up to 75% above that for growing cells and (b) a continuous decline to as little as 4%. Each yeast behaved differently. The rates did not remain constant during the periods of starvation often used for studies on the transport of sugars into yeasts. For Pichia pinus, there were striking differences, associated with starvation, between the transport of 2-deoxy-d-glucose and d-glucose, despite evidence that the two sugars enter this yeast by means of the same carrier. Some physiological explanations for these findings are discussed.
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  • 47
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    Archives of microbiology 115 (1977), S. 55-60 
    ISSN: 1432-072X
    Keywords: Malate dehydrogenase ; Inactivation ; Glucose metabolism ; Glyceraldehyde-3-phosphate ; Saccharomyces cerevisiae ; Glyoxylate cycle
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    Topics: Biology
    Notes: Abstract The cytoplasmic malate dehydrogenase in the yeast Saccharomyces cerevisiae is known to be inactivated by a glucose dependent process. In this paper it is shown that in vivo effectors of the glucose metabolism (arsenate, iodoacetate, acetaldehyde) inhibit the inactivation or change the inactivation kinetics. In vitro it was possible to inactivate the malate dehydrogenase by addition of the glucose metabolite glyceraldehyde 3-phosphate. The physiological relevance of this modification and the effect of malate dehydrogenase inactivation on the glyoxylate cycle in yeast is discussed.
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  • 48
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    Archives of microbiology 117 (1978), S. 197-201 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Glycolytic pathway ; Fermentation rate ; Protein concentration ; Kinetic parameters ; Glycolytic enzymes
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract 1. The problem of the influence of protein concentration on the kinetic parameters of enzymes has been approached studying the glycolytic enzymes from Saccharomyces cerevisiae in permeabilized cells (in situ). 2. The values of K m and V max for the different enzymes were essentially the same in dilute solutions of protein and in concentrated ones (in situ) except in the case of enolase where some differences were observed. 3. Functioning of the whole glycolytic pathway was compared in situ and in vitro measuring the rate of the fermentation of glucose. The rate of fermentation in situ was two fold higher than in vitro and the lag before active fermentation was also much shorter. 4. An unidentified phosphorylated compound, possibly polyphosphate, accumulates during the fermentation of glucose under in situ conditions.
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  • 49
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    Archives of microbiology 128 (1980), S. 157-161 
    ISSN: 1432-072X
    Keywords: Lactobacillus brevis ; Streptococcus lactis ; Saccharomyces cerevisiae ; Concanavalin A ; Symbiosis ; Tibi
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    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.
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  • 50
    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
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    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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  • 51
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Borrelidin ; Aspartokinase ; Feedback regulation ; Threonine pool ; Homoserine pool ; S-Adenosylmethionine
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    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.
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  • 52
    ISSN: 1432-072X
    Keywords: Agglutination substance ; α Pheromone ; Cell cycle ; Ethyl N-phenylcarbamate ; Mating reaction ; Microtubules ; Saccharomyces cerevisiae
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    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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  • 53
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Glucan synthetase ; EDTA ; Magnesium ; Sucrose ; Fluoride
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    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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  • 54
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    Archives of microbiology 137 (1984), S. 357-361 
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Killer toxin ; Extracellular glycoprotein
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    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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  • 55
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    Archives of microbiology 125 (1980), S. 89-95 
    ISSN: 1432-072X
    Keywords: Sulfur dioxide ; Sulfite ; Air polluting substances ; Saccharomyces cerevisiae ; Active agent of irreversible cell inactivation
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    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.
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  • 56
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    Archives of microbiology 110 (1976), S. 279-286 
    ISSN: 1432-072X
    Keywords: Thallium accumulation ; Saccharomyces cerevisiae ; Escherichia coli ; Bacillus megaterium KM ; Thallium toxicity ; Potassium ; Microbial growth
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    Notes: Abstract Thallium sulphate inhibited microbial growth, withBacillus megaterium KM, more sensitive to the metal thanSaccharomyces cerevisiae andEscherichia coli. Inhibition ofB. megaterium KM andS. cerevisiae, but not ofE. coli, was alleviated by increasing the potassium concentration of the medium; inhibition of respiration ofS. cerevisiae, but not ofE. coli, was similarly alleviated. Thallium was rapidly bound, presumably to cell surfaces, byS. cerevisiae andE. coli, and was progressively accumulated by energy-dependent transport systems (probably concerned primarily with potassium uptake) with both organisms. Thallium uptake kinetics suggested more than one transport system operated in yeast, possibly reflecting a multiplicity of potassium transport systems. ApparentK m andK i values for competitive inhibition of thallium uptake by potassium indicatedS. cerevisiae to have a higher affinity for thallium uptake than for potassium, whileE. coli had a transport system with a higher affinity for potassium than for thallium. The likely systems for thallium transport are discussed. A mutant ofE. coli with tenfold decreased sensitivity to thallium was isolated and apparently effected surface binding of thallium in amounts equivalent to the wild type organism, but showed no subsequent uptake and accumulation of the metal from buffer, even though it was able to accumulate potassium to normal intracellular concentrations during growth.
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  • 57
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    Archives of microbiology 111 (1976), S. 13-19 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Sporulation ; Ribonuclease ; Turnover of nucleic acids
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    Notes: Abstract The turnover of nucleic acids and changes in ribonuclease activity during sporulation of Saccharomyces cerevisiae were studied. In the sporulating strains, 37–58% of vegetatively synthesized RNA were degraded during the sporulation process. The degree of degradation of vegetative RNA was proportional to the sporulation ability. In the non-sporulating strains, the degradation of vegetative RNA was less than 28% in the sporulation medium. Accompanied by the degradation of vegetative RNA, a ribonuclease activity increased several times during sporulation. We have found a close relation among the sporulation rate, the degree of the degradation of vegetative RNA and the increase in ribonuclease activity in the sporulation medium, using cells of which sporulation ability was repressed by changing the age or carbon source in various degrees.
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  • 58
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    Archives of microbiology 108 (1976), S. 27-33 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Mating reaction ; Sexual cell agglutination ; α substance-I ; Agglutination factor
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    Notes: Abstract A diffusible sex-specific substance called α substance-I (αS-I) was isolated from culture filtrate of α type strains of the yeast Saccharomyces cerevisiae. The isolated αS-I, an oligopeptide, induced sexual cell agglutinability in inducible a type strains and enhanced the agglutinability in constitutive a type strains. The induction of sexual agglutinability was detected in 30 min and reached maximum in 90 min, when 0.2 μg/ml of αS-I was added to inducible a type cells. The a type-specific factor responsible for sexual cell agglutination, called a type agglutination factor (aAF), was shown to be produced during the induction or the enhancement of agglutinability of a type cells by αS-I. The aAF produced in response to αS-I was not different in the susceptibility to proteolytic enzymes and disulfide-cleaving agents from those produced constitutively in the absence of αS-I.
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  • 59
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    Archives of microbiology 114 (1977), S. 77-81 
    ISSN: 1432-072X
    Keywords: Mannoproteins ; Dolichyl monophosphate mannose ; Subcellular site of glycosylation ; Secretion ; Saccharomyces cerevisiae
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    Notes: Abstract Membranes of Saccharomyces cerevisiae were separated on urografin gradients. The specific activity of the light membranes (endoplasmic reticulum), the Golgi-like vesicles and the plasma membrane in transferring mannosyl residues from GDP-mannose to mannoproteins and to dolichyl monophosphate has been determined. The first mannose of the O-glycosidically linked manno-oligosaccharides is incorporated with the highest specific activity by the endoplasmic reticulum. The incorporation of the second to fourth mannosyl groups is catalysed with increasing activity also by the Golgi-like vesicles and the plasma membrane. The incorporation of mannosyl groups into weak alkali-stable positions (N-glycosidically linked chains) is carried out with almost the same specific activity by all three membrane fractions, however, dolicholdependent and-independent steps could not be distinguished as yet. The results are discussed in terms of a sequential addition of sugar residues along the route of export of the mannoproteins. The dolichol-dependent steps seem to occur on the endoplasmic reticulum and thus very carly in the event.
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  • 60
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    Archives of microbiology 113 (1977), S. 293-302 
    ISSN: 1432-072X
    Keywords: Aminopterin ; Saccharomyces cerevisiae ; Polyploid ; Oxidative-fermentative yeast ; Ultrastructure ; Bioassay ; Synchrony
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    Notes: Abstract In a related brewing study detailed characteristics of fermentations displaying effective yeastaminopterin interaction were presented. Fermentative yeast types (certain Saccharomyces species and Selenotila intestinalis) proved effective aminopterin reactors whereas oxidative yeasts (certain Candida, Cryptococcus, Pichia, Rhodotorula, Saccharomyces, and Trigonopsis species) proved ineffective reactors. In general effective reactors were polyploids characterized by the lack of film or pellicle formation and ineffective reactors the opposite. In stationary fermentations the Fleischmann 139 strain of S. cerevisiae proved a fair reactor. When aerated it proved an ineffective reactor and aminopterin or products there-of stimulated growth. Conversely aeration enhanced aminopterin activity of effective reactor yeasts. The positive effect of biotin on aminopterin activity and the negative effect of yeast extract, L-asparagine, adenine and thymine is shown and compared and contrasted with earlier reported studies. These findings supported by outside data suggest that oxidative yeasts (and bacteria) can readily elicit enzymes capable of inactivating aminopterin whereas fermentative types are lacking in this capability. Finally that past yeast-aminopterin studies were conducted with oxidative yeast types. Advantages of effective aminopterin reactor yeasts to be published elsewhere include improved ultrastructure using KMnO4−OsO4 fixation, a yeast bioassay procedure for detecting aminopterin in plasma and urine, and cell synchronization.
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  • 61
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    Keywords: Environment ; Kluyveromyces fragilis ; Lipids ; Saccharomyces cerevisiae ; Sterol esters ; Triacylglycerols ; Vesicles
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    Notes: Abstract Saccharomyces cerevisiae, grown aerobically or anaerobically under conditions which induce a requirement for a sterol and an unsaturated fatty acid, synthesized approximately the same amounts of neutral lipid and intracellular low-density vesicles, although the neutral lipids in aerobically-grown cells contained more esterified sterol and less triacylglycerol than those in anaerobically-grown cells. Kluyveromyces fragilis synthesized much less neutral lipid and a smaller quantity of low-density vesicles than S. cerevisiae whether grown at 30°C (generation time 1.1 h) or 20°C (generation time 2.1 h). Both yeasts synthesized highly saturated triacylglycerols, relatively unsaturated phospholipids, and esterified sterols with an intermediate degree of unsaturation irrespective of the conditions under which they were grown. Free sterols in the yeasts were rich in ergosterol and 22(24)-dehydroergosterol, while the esterified sterol fractions were richer in zymosterol.
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  • 62
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    Archives of microbiology 132 (1982), S. 141-143 
    ISSN: 1432-072X
    Keywords: Phosphoenolpyruvate carboxykinase ; Gluconeogenesis ; Saccharomyces cerevisiae ; Mutant
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    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.
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  • 63
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    Archives of microbiology 129 (1981), S. 146-149 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Inhibition of tryptophanyl-tRNA synthetase ; Mode of action of tryptophan analogues ; Tryptophan analogue degradation
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    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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  • 64
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    Keywords: Fructose-1,6-bisphosphatase ; Phosphofructokinase ; Antagonistic enzymes ; Glycolytic intermediates ; ATP ; Saccharomyces cerevisiae
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    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.
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  • 65
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    Keywords: Hansenula wingei ; Induction ; Saccharomyces cerevisiae ; Saccharomyces kluyveri ; Sex pheromone ; Sexual agglutinability ; Sexual agglutination
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    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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  • 66
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    Keywords: Saccharomyces cerevisiae ; Bud scar ; Scar ring ; Glucan ; Chitin ; Mannan ; Cytology ; Electron and X-ray diffractions ; Physico-chemical characterization
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    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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  • 67
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    Keywords: Agglutination substance ; Cell-cell recognition ; Glycoprotein ; Mating ; Saccharomyces cerevisiae ; Sexual agglutinability ; Yeast
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    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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  • 68
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    Archives of microbiology 131 (1982), S. 298-301 
    ISSN: 1432-072X
    Keywords: Yeast ; Protoplast ; Compartmentation ; Vacuole ; Trehalose ; Trehalase ; Carbohydrate metabolism ; Saccharomyces cerevisiae
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    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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  • 69
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    Archives of microbiology 133 (1982), S. 242-248 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Tryptophan degradation to tryptophol ; Degradation-defective mutant strain ; Aromatic aminotransferases ; Tryptophan accumulation
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    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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  • 70
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    Keywords: α Pheromone ; Cell cycle ; G1 arrest ; Hansenula wingei ; Saccharomyces cerevisiae ; Shmoo
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    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.
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  • 71
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    Archives of microbiology 105 (1975), S. 187-192 
    ISSN: 1432-072X
    Keywords: Peroxisomes (microbodies) ; Saccharomyces cerevisiae ; Catalase ; Urate oxidase ; Glyoxylate cycle
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    Notes: Abstract Peroxisomes were isolated from derepressed (lactose grown) Saccharomyces cerevisiae cells following homogenization with a “Merkenschlager” cell mill (at 0°C using glass beads). Catalase and urate oxidase, along with low activities of d-amino acid oxidase and l-α-hydroxyacid oxidase (glycollate oxidase), were associated with the peroxisomes. No catalase activity was present in glucose repressed cells. When protoplasts prepared from derepressed cells were used for peroxisome isolation, catalase activity was not sedimentable through gradients. Apparently peroxisomes were destroyed as the cells became fermentative during protoplast preparation. The distribution of glyoxylate cycle enzymes was examined. Isocitrate lyase was not sedimentable, suggesting that, if the enzyme is peroxisome-associated, it is either readily released or present in a labile second class of peroxisomes. Low activities of malate dehydrogenase and citrate synthetase were found in peroxisome fractions from gradients, but may represent mitochondrial contamination. Citrate synthetase was not found associated with a low-density particle as had been previously reported.
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  • 72
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    Archives of microbiology 109 (1976), S. 9-14 
    ISSN: 1432-072X
    Keywords: Candida utilis ; Saccharomyces cerevisiae ; Colloidal gold ; Cytochemistry ; Mannan ; Plasma membranes ; Scanning electron microscopy
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    Notes: Abstract The β(1→3)glucanase of Basidiomycete QM 806 was used to prepare Saccharomyces cerevisiae and Candida utilis protoplasts. Plasma membranes isolated from S. cerevisiae contained a small amount of mannose and traces of glucose and ribose. Randomly distributed α-mannan was detected by scanning electron microscopy at the surface of prefixed protoplasts using colloidal gold labelled with Concanavalin A as a marker. C. utilis protoplasts were also marked with anti-mannan antibodies. Again the distribution of mannan was random. This experiment indicated also that plasma membrane mannan has the same immunochemical determinants as cell wall mannan. It is hypothesized that mannan is mainly located in the outer layer of plasma membranes.
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  • 73
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    Archives of microbiology 109 (1976), S. 221-225 
    ISSN: 1432-072X
    Keywords: Phosphoenolpyruvate carboxykinase ; Inactivation ; Saccharomyces cerevisiae ; Carbohydrate metabolism
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    Notes: Abstract Phosphoenolpyruvate carboxykinase showed high activity in Saccharomyces cerevisiae grown on gluconeogenic carbon sources. Addition of glucose to such cultures caused a rapid loss of the phosphoenolpyruvate carboxykinase activity. Fructose or mannose had the same effect as glucose, while 2-deoxyglucose or galactose were without effect. The inactivation was an irreversible process, since the regain of the activity was dependent of de novo protein synthesis. Cycloheximide did not prevent inactivation. All strains of the genus Saccharomyces tested showed inactivation of their phosphoenolpyruvate carboxykinase upon addition of glucose; this behaviour was not restricted to this genus.
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  • 74
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    Archives of microbiology 107 (1976), S. 229-231 
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    Keywords: Saccharomyces cerevisiae ; pH ; Sulphite formation
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    Notes: Abstract The influence of the initial pH of the substrate on the sulphite formation of three low-sulphite-and five high-sulphite-forming yeasts is described. Four distinctly different groups become apparent. The need for better evaluation of pure culture wine yeasts is stressed.
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  • 75
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    Archives of microbiology 113 (1977), S. 159-161 
    ISSN: 1432-072X
    Keywords: Baker's yeast ; Spheroplasts ; Gluconeogenesis ; Location ; Density gradient centrifugation ; Saccharomyces cerevisiae
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    Notes: Abstract The subcellular location of hexose diphosphatase, phosphoenolpyruvate carboxykinase and pyruvate carboxylase in baker's yeast (Saccharomyces cerevisiae) was investigated by density gradient centrifugation of spheroplast lysates obtained by osmotic shock treatment of spheroplasts and centrifugation for 10000 g x min. On the evidence obtained from zonal separations these three enzymes of gluconeogenesis are most probably located in the soluble cytosol.
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  • 76
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    Archives of microbiology 114 (1977), S. 287-288 
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    Keywords: Saccharomyces cerevisiae ; Mating reaction ; Sexual agglutination ; Temperature dependency
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    Notes: Abstract Temperature dependency of sexual agglutination in Saccharomyces cerevisiae was found. Of 31 strains tested, which showed normal agglutination when cultured at 25°C, 29 strains lost their sexual agglutinability when they were grown at 37°C.
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  • 77
    ISSN: 1432-072X
    Keywords: Yeast flocculation ; Chemical modification of cell surface components ; Floc-forming ability ; Brewer's yeast ; Saccharomyces cerevisiae ; Deflocculation
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    Notes: Abstract Effects of treatments with proteolytic enzymes and protein-modifying reagents on flocculation of brewer's yeast IFO 2018 were investigated. The floc-forming ability of the yeast cells was irreversibly eliminated by treatment with papain, trypsin, chymotrypsin or pepsin, indicating that certain proteins on the cell surface participate in the yeast flocculation. Chemical modification with reagents, known to act on disulfide bridges, carboxyl and/or phosphate groups, phenolic groups, amino groups, and imidazole groups, also destroyed the ability to flocculate, although in some cases a high concentration (8 M) of urea was necessary in addition to protein-modifying reagents. Thus, it is suggested strongly that these functional groups of amino acid residues of the proteins are essential for the floc-forming ability of brewer's yeast cells.
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  • 78
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    Archives of microbiology 117 (1978), S. 239-245 
    ISSN: 1432-072X
    Keywords: Plasma membrane ; Lipids ; Saccharomyces cerevisiae ; Ethanol tolerance ; Sterols ; Fatty-acyl residues
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    Topics: Biology
    Notes: Abstract Populations of cells suspended anaerobically in buffered (pH 4.5) M ethanol remained viable to a greater extent when their plasma membranes were enriched in linoleyl rather than oleyl residues irrespective of the nature of the sterol enrichment. However, populations with membranes enriched in ergosterol or stigmasterol and linoleyl residues were more resistant to ethanol than populations enriched in campesterol or cholesterol and linoleyl residues. Populations enriched in ergosterol and cetoleic acid lost viability at about the same rate as those enriched in oleyl residues, while populations grown in the presence of this sterol and palmitoleic acid were more resistant to ethanol. Suspending cells in buffered ethanol for up to 24 h did not lower the ethanol concentration.
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  • 79
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    Keywords: Ethanol inhibition ; Solute accumulation ; Saccharomyces cerevisiae ; Plasma membrane ; Lipids
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    Notes: Abstract Incorporation of ethanol (1.0 or 1.25 M) into exponential-phase cultures of Saccharomyces cerevisiae NCYC 366 growing anaerobically in a medium supplemented with ergosterol and an unsaturated fatty acid caused a retardation in growth rate, which was greater when the medium contained oleic rather than linoleic acid. Ethanol incorporation led to an immediate drop in growth rate, and ethanol-containing cultures grew at the slower rate for at least 10 h. Incorporation of ethanol (0.5 M) into buffered (pH 4.5) cell suspensions containing d-[6-3H] glucose, d-[1-14C] glucosamine, l-[U-14C] lysine or arginine, or KH2 32PO4 lowered the rate of solute accumulation by cells. Rates of accumulation of glucose, lysine and arginine were retarded to a greater extent when cells had been grown in the presence of oleic rather than linoleic acid. This difference was not observed with accumulation of phosphate. Ethanol was extracted from exponential-phase cells by four different methods. Cells grown in the presence of linoleic acid contained a slightly, but consistently, lower concentration of ethanol than cells grown in oleic acid-containing medium. The ethanol concentration in cells was 5–7 times greater than that in the cell-free medium.
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  • 80
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Catabolite repression and inactivation ; Inhibition of protease B
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    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.
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  • 81
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    Archives of microbiology 132 (1982), S. 236-240 
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    Keywords: α Pheromone ; Cycloheximide ; Inducible a strain ; Phenylmethylsulfonyl fluoride ; Saccharomyces cerevisiae ; Sexual agglutinability ; Temperature-sensitive
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    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.
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  • 82
    ISSN: 1432-072X
    Keywords: Candida utilis ; Saccharomyces cerevisiae ; Zygosaccharomyces bailii ; Compartmentation ; Vacuoles ; Internal pH ; Phosphate ; Glycolysis ; Nuclear magnetic resonance
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    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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  • 83
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    Archives of microbiology 134 (1983), S. 64-67 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Anaerobic growth ; Hungate technique ; Tween 80 ; Ergosterol
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    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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  • 84
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    Archives of microbiology 134 (1983), S. 171-174 
    ISSN: 1432-072X
    Keywords: Acetate growth medium ; Anti-microtubule agent ; Bud initiation ; Ethyl N-phenylcarbamate ; Meiosis ; Mitotic cell cycle ; Saccharomyces cerevisiae ; Sporulation induction
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    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.
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  • 85
    ISSN: 1432-072X
    Keywords: Nucleotide pools ; Continuous cultivation ; Synchronized growth ; Saccharomyces cerevisiae
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    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.
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  • 86
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    Archives of microbiology 104 (1975), S. 23-28 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Yeast ; Chemostat ; Nutrient Concentration ; Thermal Death ; Thermal Association ; Optimum Temperature for Growth ; Maximum Temperature for Growth ; Microbial Ecology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Saccharomyces cerevisiae was grown in a chemostat under glucose limitation at three superoptimal temperatures. In each steady state the specific growth rate was the sum of the dilution rate and the specific death rate, exponential death concurring with exponential growth. The specific death rate was a function of the temperature while the specific growth rate was a function of both the temperature and the concentration of the limiting nutrient. Each superoptimal temperature was characterized by a critical glucose concentration below which net growth was not possible. The critical glucose concentration increased with the temperature. Consequently the maximum temperature for growth was a function of the concentration of the limiting nutrient and approached the optimum temperature for growth with decreasing glucose concentrations.
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  • 87
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    Archives of microbiology 106 (1975), S. 271-273 
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    Keywords: Saccharomyces cerevisiae ; Baker's yeast ; Gluconeogenetic enzymes ; Chemostat ; Oxygen
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    Notes: Abstract 1. The effect of aeration on the key enzymes of gluconeogenesis was studied in baker's yeast (Saccharomyces cerevisiae) and in a nonrespiratory variant of S. cerevisiae grown under glucose limitation. 2. In baker's yeast phosphoenolpyruvate carboxykinase, hexosediphosphatase and isocitrate lyase were completely repressed under anaerobic conditions. Their repression could be partially reversed by using intense aeration. 3. In the nonrespiratory variant these enzymes were absent independently of aeration. 4. Pyruvate carboxylase of baker's yeast showed maximal activity under anaerobic conditions. In the nonrespiratory variant pyruvate carboxylase had low activity under both anaerobic and aerobic conditions.
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  • 88
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    Archives of microbiology 108 (1976), S. 149-152 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Sporulation ; Ribonuclease ; Caffeine ; Cycloheximide
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    Notes: Abstract Changes in RNase activity during sporulation of a homothallic diploid strain of Saccharomyces cerevisiae were measured in caffeine-treated and non-treated cells. 1. In caffeine-treated cells soon after the transfer to the sporulation medium a significant increase in RNase activity was observed; in control cells the rise of RNase activity was less and started after a lag period of 5 h. The final activity of RNase was about twice as high in caffeine-treated cells as in control cells. 2. Increase in RNase activity during sporulation was sensitive to cycloheximide in control cells, but insensitive in caffeine-treated cells. 3. RNases from vegetative cells and from sporulating ones are different in their K m values. Relation of the changes in RNase activity to premeiotic DNA synthesis is discussed.
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  • 89
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    Archives of microbiology 108 (1976), S. 293-298 
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Maximum temperature for growth ; Thermal death ; Linear thermodynamic compensation ; Non-linear thermodynamic compensation ; Isokinetic temperature
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    Topics: Biology
    Notes: Abstract Sixty eight Arrhenius plots of thermal death in six mesophilic yeast species, tested at various concentrations of NaCl, lacked an isokinetic temperature. Nevertheless the ΔH #/ΔS # plot was apparently linear with a slope corresponding to 314° K. It was concluded that linear thermodynamic compensation of thermal death is non-existent in heterogeneous groups of yeasts and is unlikely to occur in heterogeneous groups of other organisms and that ΔH #/ΔS # plots lack sensitivity for the detection of non-linearity over narrow temperature ranges. However, the ΔH # and ΔS # parameters of thermal death displayed non-linear compensation in such a way that the extrapolated Arrhenius plots of death attained nearly identical values near the respective maximum temperatures for growth. Linear thermodynamic compensation occurred in each of the six strains, when stationary populations of the same strain were tested at various NaCl concentrations. On the other hand, exponential populations of each of the strains, tested in the same way, lacked an isokinetic temperature of thermal death. The significance of linear and non-linear thermodynamic compensation in biological rate processes is discussed.
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  • 90
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    Archives of microbiology 110 (1976), S. 313-318 
    ISSN: 1432-072X
    Keywords: Yeast protoplasts ; Saccharomyces cerevisiae ; Conjugation ; Cell wall ; Morphogenesis
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    Topics: Biology
    Notes: Abstract Protoplasts prepared from complementary haploid strains ofSaccharomyces cerevisiae were studied with regard to their ability of conjugating. Neither fresh protoplasts nor the growing protoplasts possessing fibrillar walls exhibited sex specific agglutination or fusion. However, they were capable of inducing sexual activation in normal cells of opposite mating type. After completing the regeneration of cell walls the protoplasts could conjugate either with each other or with cells of opposite sex. The frequency of conjugations was low, about 1%, and was largely dependent on the degree of completition of the wall during regeneration. From the results the following conclusions may be drawn: 1. The initiation of mating is dependent on the integrity of the cell wall. 2. The sex specific morphogenetic changes do not occur in wall-less protoplasts but may happen after the protoplasts have regenerated their cell walls. 3. The lysis of cell walls does not occur until the walls come into close contact. 4. The fusion of plasma membranes in sex-activated protoplasts cannot be induced by artefucial agglutination.
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  • 91
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    Archives of microbiology 113 (1977), S. 303-307 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Specific growth rate ; Growth control ; Glucose transfer ; Glucose-6-phosphate ; Maintenance requirements
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    Topics: Biology
    Notes: Abstract The specific growth rate (μ) of a respiration-deficient mutant of Saccharomyces cerevisiae growing under defined experimental conditions in batch culture (mineral medium plus glucose and vitamins at 25°C) varied from experiment to experiment over a wide range (0.10–0.24 h-1) and showed a normal distribution. Neither the age of the culture, the history of the inoculum, nor experimental error accounted wholy for the variability of μ. The variation was positively correlated with the specific rate of glucose transfer and negatively with the specific rate of production of non-fermentative CO2. The yield decreased with μ implying higher maintenance requirements in batch culture (4.7 mmoles g-1 h-1) than in continuous culture (0.8 mmoles g-1 h-1). It was concluded that the strain is capable of establishing any one of several steady states of growth under the same experimental conditions, each steady state displaying some buildin inertia with respect to change. The variations of the specific rates of glucose transfer and non-fermentative CO2 production, and of the yield appeared to be consequences rather than causes of the variation of μ. The ultimate causes of the variation of μ remained unidentified.
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  • 92
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    Archives of microbiology 114 (1977), S. 91-92 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Cell wall ; Glucan ; Mannan ; Synchronous culture
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    Topics: Biology
    Notes: Abstract The mode of increase in cell wall polysaccharides of yeast (glucan and mannan) during cell cycle was analyzed using cell wall samples obtained from a synchronous culture. The increase in mannan and total glucan proceeded almost linearly throughout the cell cycle except for a short period of their leveling off at the time of cell division. However, the constituents of glucan behaved characteristically: Alkalisoluble glucan and insoluble glucan increased mainly in the former and the latter half of the cell cycle, respectively.
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  • 93
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    Archives of microbiology 115 (1977), S. 307-316 
    ISSN: 1432-072X
    Keywords: Anthranilate synthase, feedback inhibition of ; Saccharomyces cerevisiae ; Tryptophan analogues, mode of action of ; Tryptophan biosynthetic enzymes ; Tryptophan pool
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In an analysis of the effects of various tryptophan and indole analogues in Saccharomyces cerevisiae we determined the mechanisms by which they cause growth inhibition: 4-Methyltryptophan causes a reduction in protein synthesis and a derepression of the tryptophan enzymes despite of the presence of high internal levels of tryptophan. This inhibition can only be observed in a mutant with increased permeability to the analogue. These results are consistent with but do not prove an interference of this analogue with the charging of tryptophan onto tRNA. 5-Methyltryptophan causes false feedback inhibition of anthranilate synthase, the first enzyme of the tryptophan pathway. This inhibits the further synthesis of tryptophan and results in results in tryptophan limitation, growth inhibition and derepression of the enzymes. Derepression eventually allows wild type cells to partially overcome the inhibitory effect of the analogue. 5-Fluoroindole is converted endogenously to 5-fluorotryptophan by tryptophan synthase. Both endogenous and externally supplied 5-fluorotryptophan are incorporated into protein. This leads to intoxication of the cells due to the accumulation of faulty proteins. 5-Fluorotryptophan also causes feedback inhibition of anthranilate synthase and reduces the synthesis of tryptophan which would otherwise compete with the analogues in the charging reaction. Indole acrylic acid inhibits the conversion of indole to tryptophan by tryptophan synthase. This results in a depletion of the tryptophan pool which, in turn, causes growth inhibition and derepression of the tryptophan enzymes.
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  • 94
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    Archives of microbiology 117 (1978), S. 73-77 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Ascospores ; Germination
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    Topics: Biology
    Notes: Abstract The wall of mature ascospores ofSaccharomyces cerevisiae showed in sections under the electron microscope a dark outer layer and a lighter inner layer. The latter was composed of a greyish inner part and a light outer part. During germination, the spore grew out at one side and the dark outer layer was broken. Of the light inner layer, the inner greyish part became the wall of the vegetative cell, but the extented part of the cell had a new wall.
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  • 95
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    Archives of microbiology 117 (1978), S. 269-276 
    ISSN: 1432-072X
    Keywords: Succinic acid ; Fermentation ; Saccharomyces cerevisiae ; α-ketoglutarate dehydrogenase
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    Topics: Biology
    Notes: Abstract 1. Succinic acid is formed in amounts of 0.2–1.7 g/l by fermenting yeasts of the genusSaccharomyces during the exponential growth phase. No differences were observed between the various species, respiratory deficient mutants and wild type strains. 2. At low glucose concentrations the formation of succinic acid depended on the amount of sugar fermented. However, the nitrogen source was found to be of greater importance than the carbon source. 3. Of all nitrogen sources, glutamate yielded the highest amounts of succinic acid. Glutamate led to an oxidative and aspartate to a reductive formation of succinic acid. 4. A reductive formation of succinic acid by the citric acid cycle enzymes was observed with malate. This was partially inhibited by malonate. No evidence was obtained that the glyoxylate cycle is involved in succinic acid formation by yeasts. 5. Anaerobically grown cells ofSaccharomyces cerevisiae contained α-ketoglutarate dehydrogenase. Its activity was found in the 175000 x g sediment after fractionated centrifugation. The specific activity increased 6-fold after growth on glutamate as compared with cells grown on ammonium sulfate. 6. The specific activities of malate dehydrogenase, fumarase, succinate dehydrogenase, succinylcoenzymeA synthetase, α-ketoglutarate dehydrogenase and glutamate dehydrogenase (nicotinamide adenine dinucleotide dependent) were determined in yeast cells grown on glutamate or ammonium sulfate. Similar results were obtained with a wild type strain and a respiratory deficient mutant. The latter did not contain succinate dehydrogenase. 7. In fermenting yeasts succinic acid is mainly formed from glutamate by oxidation.
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  • 96
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    Archives of microbiology 119 (1978), S. 213-214 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Cell wall ; Glucan ; Mannan ; Cell cycle
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    Topics: Biology
    Notes: Abstract Reevaluation and comparison of seemingly contradictory literature data on the mode of synthesis of wall polysaccharides during the cell cycle ofSaccharomyces cerevisiae explained the source of discrepancies and demonstrated their general consonance in the following points: 1. The rate of synthesis of glucan and mannan is not constant and does not increase continuously throughout the entire cell cycle. 2. The rate of synthesis of both polysaccharides is considerably reduced at the time of cell division and in the prebudding phase.
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  • 97
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    Archives of microbiology 115 (1977), S. 1-7 
    ISSN: 1432-072X
    Keywords: Cell walls ; Chitin ; Colloidal gold ; Concanavalin A ; Cytochemistry ; Mannan ; Wheat germ agglutinin ; Yeast ; Saccharomyces cerevisiae ; Candida utilis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mannan was located on thin sections of Saccharomyces cerevisiae and Candida utilis with the homologous anti-mannan antibodies or with Concanavalin A, both labelled with gold granules. Fully synthesized mannan was found in the cell walls, on the plasmalemma and within the cytoplasm sometimes associated with vesicles and vacuoles. Chitin or its oligomers were located with wheat germ agglutinin in the bud scars but also in the cell wall and the cytoplasm near the plasmalemma. Both mannan and chitin or its oligomers were found in the forming septum and are synthesized within the cytoplasm. The gold method was also suitable for marking mannan and chitin simultaneously.
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  • 98
    ISSN: 1432-072X
    Keywords: Sulfur dioxide ; Sulfite ; Air polluting substances ; Saccharomyces cerevisiae ; ATP hydrolysis ; Reversibility of sulfite effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sulfite, at concentrations above 1 mM and at a pH below 4, caused cell death in Saccharomyces cerevisiae X2180 as measured by the colony-forming capacity. A rapid decrease in the ATP content was observed prior to cellular death. The depletion of ATP was reversible and the lethal effect could be prevented if the cells were exposed to sulfite for periods of less than 1 h. Extent and rate of ATP depletion were dependent on time, pH value, temperature and sulfite concentrations.
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  • 99
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    Archives of microbiology 124 (1980), S. 285-287 
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Heat killing ; Membrane damage ; Genetic damage ; Growth temperature
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    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.
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  • 100
    ISSN: 1432-072X
    Keywords: Glycoprotein ; Inducible strains ; Saccharomyces cerevisiae ; Sexual agglutinability ; Tunicamycin
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    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.
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