Summary
Mutant strains of Saccharomyces cerevisiae auxotrophic for deoxythymidine monophosphate (dTMP) were isolated and characterized. Two distinct classes of auxotrophs were obtained. One class had a simple requirement for dTMP and was analogous to thymine-requiring bacteria. The second class required dTMP, adenine, histidine and methionine and this complex nutritional phenotype was due to defects in folate metabolism. The dTMP-dependent growth of respiratory-competent grande auxotrophs was found to be markedly affected by media composition and carbon source. In the absence of dTMP thymineless death occurred in both mutant classes.
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Barclay, B.J., Little, J.G.: Genetic damage during thymidylate starvation in Saccharomyces cerevisiae. Mol. Gen. Genet. 160, 33–40 (1978)
Bassett, R., Weir, D.G., Scott, J.M.: The identification of hexahepta-and octaglutamates as the polyglutamyl form of folate found throughout the growth cycle of yeast. J. Gen. Microbiol. 93, 169–172 (1976)
Bisson, L., Thorner, J.: Thymidine 5′-monophosphate — requiring mutants of Saccharomyces cerevisiae are deficient in thymidylate synthetase. J. Bacteriol. 132, 44–50 (1977)
Blakley, R.L.: The biochemistry of folic acid and related pteridines Amsterdam, North-Holland 1969
Brendel, M.: A simple method for the isolation and characterization of thymidylate uptaking mutants in Saccharomyces cerevisiae. Mol. Gen. Genet. 147, 209–215 (1976)
Brendel, M., Fäth, W.W.: Isolation and characterization of mutants of Saccharomyces cerevisiae auxotrophic and conditionally auxotrophic for 5′-dTMP. Z. Naturforsch. 29c, 733–738 (1974)
Brendel, M., Haynes, R.H.: Interactions among genes controlling sensitivity to radiation and alkylation in yeast. Mol. Gen. Genet. 125, 197–216 (1973)
Carter, B.L.A.: The organization and replication of genetic material in yeast. Cell 6, 259–268 (1975)
Chung, S.T., Greenberg, G.R.: Temperature-sensitive revertants. Biochem. Biophys. Res. Commun. 45, 1337–1343 (1971)
Cohen, S.S., Barner, H.D.: Studies on unbalanced growth in Escherichia coli. Proc. Nat. Acad. Sci. U.S.A. 40, 885–893 (1954)
Cox, B., Game, J.: Repair systems in Saccharomyces. Mutat. Res. 26, 257–264 (1974)
Fäth, W.W., Brendel, M., Laskowski, W., Lehmann-Brauns, E.: Economizing DNA-specific labelling by deoxythymidine-5′-monophosphate in Saccharomyces cerevisiae. Mol. Gen. Genet. 132, 335–345 (1974)
Game, J.C.: Yeast cell cycle mutant cdc 21 is a temperature-sensitive thymidylate auxotroph. Mol. Gen. Genet. 146, 313–315 (1976)
Game, J.C., Cox, B.S.: Synergistic interactions between rad mutations in yeast. Mutat. Res. 20, 35–44 (1973)
Grivell, A.R., Jackson, J.F.: Thymidine kinase: evidence for its absence from Neurospora crassa and some other microorganisms, and the relevance of this to the specific labelling of deoxyribonucleic acid. J. gen. Microbiol. 54, 307–317 (1968)
Hartwell, L.H.: Three additional genes required for deoxyribonucleic acid synthesis in Saccharomyces cerevisiae. J. Bacteriol. 115, 966–974 (1973)
Haynes, R.H.: DNA repair in yeast. In: Molecular mechanisms for repair of DNA. Part B (P.C. Hanawalt and R.B. Setlow, eds.), pp. 529–540. New York: Plenum Publishing 1975
Hoffbrand, A.V.: Synthesis and breakdown of natural folates (folate polyglutamates). Prog. Hematol. 9, 85–105 (1975)
Jannsen, S., Witte, I., Megnet, R.: Mutants for the specific labelling of DNA in Saccharomyces cerevisiae. Biochim. Biophys. Acta 299, 681–685 (1973)
Lam, K.B., Jones, E.W.: Mutations affecting levels of tetrahydrofolate interconversion enzymes in Saccharomyces cerevisiae. I. Enzyme levels in ade 3–41 and ADE 15, a dominant adenine auxotroph. Mol. Gen. Genet. 123, 199–208 (1973)
Laskowski, W., Lehmann-Brauns, E.: Mutants of Saccharomyces able to grow after inhibition of thymidine phosphate synthesis. Mol. Gen. Genet. 125, 275–277 (1973)
Little, J.G., Haynes, R.H.: DNA-specific labelling in yeast mutants. Genetics 74, S161 (1973)
Lomax, C.A., Gross, T.S., Woods, R.A.: New mutant types at the ade 3 locus of Saccharomyces cerevisiae. J. Bacteriol. 107, 1–7 (1971)
Luzzati, M.: Isolation and properties of a thymidylate-less mutant in Saccharomyces cerevisiae. Eur. J. Biochem. 56, 533–538 (1975)
Mahler, H.R.: Biogenetic autonomy of mitochondria. CRC Crit. Rev. Biochem. 1, 381–460 (1973)
McBurney, M.W., Whitmore, G.F.: Isolation and biochemical characterization of folate deficient mutants of chinese hamster cells. Cell 2, 173–182 (1974)
McKenzie, K.Q., Jones, E.W.: Mutants of the formyltetrahydrofolate interconversion pathway of Saccharomyces cerevisiae. Genetics 86, 85–102 (1977)
Roman, H.: A system selective for mutations affecting the synthesis of adenine in yeast. Compt. Rend. Trav. Lab. Carlsberg Physiol. 26, 299–314 (1956)
Sherman, F., Fink, G.R., Lawrence, C.W. (eds.): Methods in yeast genetics, p. 5. New York: Cold Spring Harbor Laboratory 1974
Slater, M.L.: Effect of reversible inhibition of deoxyribonucleic acid synthesis on the yeast cell cycle. J. Bacteriol. 113, 263–270 (1973)
Smith, A.E., Marcker, K.A.: N-formylmethionyl transfer RNA in mitochondria from yeast and rat liver. J. Mol. Biol. 38, 241–243 (1968)
Stone, A.B., Wilkie, D.: Cellular and mitochondrial effects of folate antagonism by pyrimethamine in Saccharomyces cerevisiae. J. Gen. Microbiol. 83, 283–293 (1974)
Toh-E, A., Ueda, Y., Kakimoto, S.I., Oshima, Y.: Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae. J. Bacteriol. 113, 727–738 (1973)
Wickner, R.B.: Mutants of Saccharomyces cerevisiae that incorporate deoxythymidine-5′-monophosphate into deoxyribonucleic acid in vivo. J. Bacteriol. 117, 252–260 (1974)
Williamson, D.H., Maroudas, N.G., Wilkie, D.: Induction of the cytoplasmic petite mutation in Saccharomyces cerevisiae by the antibacterial antibiotics erythromycin and chloramphenicol. Mol. Gen. Genet. 111, 209–223 (1971)
Wintersberger, U., Hirsch, J.: Induction of cytoplasmic respiratory deficient mutants in yeast by the folic acid analogue, methotrexate. Mol. Gen. Genet. 126, 61–70 (1973)
Zelikson, R., Luzzati, M.: Two forms of serine transhydroxymethylase, one absent in a thymidylate-less mutant in Saccharomyces cerevisiae. Eur. J. Biochem. 64, 7–13 (1973)
Zelikson, R., Luzzati, M.: Mitochondrial and cytoplasmic distribution in Saccharomyces cerevisiae of enzymes involved in folate-coenzyme-mediated one-carbon-group transfer. Eur. J. Biochem. 79, 285–292 (1977)
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Communicated by G. O'Donovan
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Little, J.G., Haynes, R.H. Isolation and characterization of yeast mutants auxotrophic for 2′-deoxythymidine 5′-monophosphate. Molec. Gen. Genet. 168, 141–151 (1979). https://doi.org/10.1007/BF00431440
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DOI: https://doi.org/10.1007/BF00431440