Contrôle biochimique de la récombinaison chez Saccharomyces cerevisiae

Luzzati, Mario; Clavilier, Léa; Péré‐Aubert, Geneviève A.; Pp, Slonimski

doi:10.1007/bf00267787

Cited by 6 publications

(1 citation statement)

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“…Natural. Medium YPG [6], containing 1 % yeast extract, 1 % Bactopeptone and 2 % glucose, was used for most of the strains. Medium YGT containing yeast extract (Difco) 1 %; glucose 2%, KH2P0, 0.1 %, MgSO, 0.05 %, dTMP 60 mg/l, was used for maintenance and growth of thymidylate-less mutants.…”

Section: Mediamentioning

confidence: 99%

Isolation and Properties of a Thymidylate-less Mutant in Saccharomyces cerevisiae

Luzzati

1975

Eur J Biochem

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A mutant, tmp3, has been isolated in Saccharomyces cerevisiae. Genetic and physiological analysis show that a single mendelian gene controls the multiple requirements for thymidylate, methionine, adenine and histidine and a neutral cytoplasmic petite character. Crude extracts of this mutant present a 60 % decrease of serine transhydroxymethylase specific activity as compared to a wild-type strain.ade3 mutants have been shown to be deficient in formyltetrahydrofolate synthetase, methenyl tetrahydrofolate cyclohydrolase and lack also one of the two forms of methylene tetrahydrofolate dehydrogenase [l-31. These deficiencies lead to the accumulation of fl,N''-methylene-tetrahydrofolate (Luzzati, M., unpublished results).In order to get a better insight into tetrahydrofolate derivatives metabolism, and since N5,N'o-methylenetetrahydrofolate is also generated in the cells by the enzymic interconversion of serine to glycine, we looked for a mutant affected in this function.A mutant, met5, lacking serine transhydroxymethylase activity was described some years ago [4], but these results have not been confirmed later (Parks, personal communication).The mutant described in this report is affected in serine transhydroxymethylase activity and presents many interesting features. MATERIALS AND METHODS Yeast StrainsHaploid strains of Saccharomyces cerevisiae were used for the initial isolation of the mutant and for ; CREROR represents resistance to chloramphenicol, erythromycin and oligomycin. All the other strains were constructed in this laboratory. MediaNatural. Medium YPG [6], containing 1 % yeast extract, 1 % Bactopeptone and 2 % glucose, was used for most of the strains. Medium YGT containing yeast extract (Difco) 1 %; glucose 2%, KH2P0, 0.1 %, MgSO, 0.05 %, dTMP 60 mg/l, was used for maintenance and growth of thymidylate-less mutants. N, [7], glycerol medium, with or without antibiotics added, was used for screening for drug resistant clones.Synthetic. 0.67 % yeast nitrogen base (Difco); different supplements were added when needed. Genetic MethodsTetrad analysis was performed by classical methods [S]. Transmission of mitochondria1 markers was studied in the usual way [9]. Cultures Techniques and Disruption of CellsUsually 1 1 of medium YGT was inoculated and the culture incubated at 28 "C with constant agitation. Cells harvested in stationary phase were washed twice with 0.05 M phosphate buffer, pH 7.5. PrepEur. J. Biochem. 56 (1975)

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Section: Mediamentioning

confidence: 99%