Summary
Mutants with defective carbon catabolite repression have been isolated in the yeast Saccharomyces cerevisiae using a selective procedure. This was based on the fact that invertase is a glucose repressible cell wall enzyme which slowly hydrolyses raffinose to yield fructose and that the inhibitory effects of 2-deoxyglucose can be counteracted by fructose. Repressed cells were plated on a raffinose-2-deoxyglucose medium and the resistant cells growing up into colonies were tested for glucose non-repressible invertase and maltase. The yield of regulatory mutants was very high. All were equally derepressed for invertase and maltase, no mutants were obtained with only non-repressible invertase synthesis which was the selected function. A total of 61 mutants isolated in different strains were allele tested and could be attributed to three genes. They were all recessive. Mutants in one gene had reduced hexokinase activities, the other class, located in a centromere linked gene, had elevated hexokinase levels and was inhibited by maltose. Mutants in a third gene were isolated on a 2-deoxyglucose galactose medium and had normal hexokinase levels. A partial derepression was observed for malate dehydrogenase in all mutants. Isocitrate lyase, however, was still fully repressible.
Similar content being viewed by others
References
Arst, H.N., Jr., MacDonald, D.W.: A mutant of Aspergillus nidulans lacking NADP-linked glutamate dehydrogenase. Molec. gen. Genet. 122, 261–265 (1973)
Barnett, J.A.: The utilization of sugars by yeasts. Advanc. Carbohydrate Chem. and Biochem. 32, 126–234 (1976)
Ciriacy, M.: Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. I. Isolation and genetic analysis of adh mutants. Mutation Res. 29, 315–326 (1975)
Ciriacy, M.: Cis-dominant regulatory mutations affecting the formation of glucose-repressible alcohol dehydrogenase (ADHH) in Saccharomyces cerevisiae. Molec. gen. Genet. 145, 327–333 (1976)
Dubois, E., Grenson, M., Wiame, J.-M.: The participation of the anabolic glutamate dehydrogenase in the nitrogen catabolite repression of arginase in Saccharomyces cerevisiae. Europ. J. Biochem. 48, 603–616 (1974)
Fincham, J.R.S.: Genetic complementation. New York and Amsterdam: W.A. Benjamin, Inc. 1966
Hackel, R.A.: Genetic control of invertase formation in Saccharomyces cerevisiae. I. Isolation and characterization of mutants affecting sucrose utilization. Molec. gen. Genet. 140, 361–370 (1975)
Haußmann, P., Zimmermann, F.K.: The role of mitochondria in carbon catabolite repression in yeast. Molec. gen. Genet. 148, 205–211 (1976)
Holzer, H.: Catabolite inactivation in yeast. Trends in Biochem. Sci. 1, 178–181 (1976)
Kinghorn, J.R., Pateman, J.A.: NAD and NADP L-glutamate dehydrogenase activity and ammonium regulation in Aspergillus nidulans. J. gen. Microbiol. 78, 39–46 (1973)
Lampen, J.O.: External enzymes of yeast: their nature and formation. Antonie v. Leeuwenhoek 34, 1–18 (1968)
Magasanik, B.: Catabolite repression. Cold Spr. Harb. Symp. quant. Biol. 26, 249–262 (1961)
Maitra, P.K.: A glucokinase from Saccharomyces cerevisiae. J. biol. Chem. 245, 2423–2431 (1970)
Maitra, P.K., Lobo, Z.: Control of glycolytic enzyme synthesis in yeast by products of the hexokinase reaction. J. biol. Chem. 246, 489–499 (1971)
Montenecourt, B.S., Kuo, S.C., Lampen, J.O.: Saccharomyces mutants with invertase formation resistant to repression by hexoses. J. Bact. 114, 233–238 (1973)
Mortimer, R.K., Hawthorne, D.C.: Yeast genetics In: A.H. Rose and J.S. Harrison (eds.), The yeasts. Vol. 1, pp. 385–460. New York: Academic Press 1969
Polakis, E.S., Bartley, W.: Changes in enzyme activities of Saccharomyces cerevisiae during growth on different carbon sources. Biochem. J. 97, 284–297 (1965)
Polakis, E.S., Bartley, W., Meek, G.A.: Changes in the activities of respiratory enzymes during aerobic growth of yeast on different carbon sources. Biochem. J. 97, 298–302 (1965)
Poll, K.W. van de: Ammonium repression in a nutant of Saccharomyces carlsbergensis lacking NADP dependent glutamate dehydrogenase activity. FEBS Letters 32 265–266 (1973)
Poll, K.W. van de, Kerkenaar, A., Schamhart, D.H.J.: Isolation of a regulatory mutant of fructose-1,6-diphosphatase in Saccharomyces carlsbergensis. J. Bact. 117, 965–970 (1974)
Rytka, J., Sledziewski, A., Litwinska, J., Bilinski, T.: Haemoprotein formation in yeast. II. Isolation of catalase regulatory mutants. Molec. gen. Genet. 145, 37–42 (1976)
Schamhart, D.H.J., Berge, A.M.A., ten, Poll, K.W. van de: A spontaneous catabolite repression insensitive mutant in Saccharomyces. J. Bact. 121, 747–760 (1975)
Witt, I., Kronau, R., Holzer, H.: Repression von Alkoholdehydrogenase, Malatdehydrogenase, Isocitratlyase und Malatsynthase in Hefe durch Glucose. Biochim. biophys. Acta (Amst.) 118, 522–537 (1966)
Zamenhof, S. In: Methods in enzymology (S.P. Colowick and N.O. Kaplan, eds.), Vol. 3, p. 702. New York: Academic Press 1957
Zimmermann, F.K.: Procedures used in the induction of mitotic recombination and mutation in the yeast Saccharomyces cerevisiae. Mutation Res. 31, 71–86 (1975)
Zimmermann, F.K., Eaton, N.R.: Genetics of induction and catabolite repression of maltase synthesis in Saccharomyces cerevisiae. Molec. gen. Genet. 134, 261–272 (1974)
Zimmermann, F.K., Kaufmann, I., Rasenberger, H., Haußmann, P.: Genetics of carbon catabolite repression in Saccharomyces cerevisiae: Genes involved in the derepression process. Molec. gen. Genet. 151, 95–103 (1977)
Author information
Authors and Affiliations
Additional information
Communicated by H. Böhme
Rights and permissions
About this article
Cite this article
Zimmermann, F.K., Scheel, I. Mutants of Saccharomyces cerevisiae resistant to carbon catabolite repression. Molec. Gen. Genet. 154, 75–82 (1977). https://doi.org/10.1007/BF00265579
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00265579