Skip to main content
Log in

Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae

Isolation and characterization of the regulatory gene GAL4

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The GAL4 gene positively regulating the expression of the gene cluster GAL7-GAL10-GAL1 in the yeast Saccharomyces cerevisiae was isolated for its ability to suppress a recessive mutation in that gene. When the isolated gene was incorporated into a multi-copy plasmid, the GAL cluster genes in the host chromosome partially escaped the normal control; a yeast that harbors the plasmid bearing the GAL4 gene synthesized the galactose-metabolizing enzymes encoded by the GAL cluster genes at a low but significant level in the absence of galactose. If the GAL7 gene was amplified along with GAL4 on the multi-copy plasmid, the constitutive synthesis of Gal-1-P uridylyl transferase encoded by GAL7 was further pronounced and the enzyme activity reached the level of the fully induced wild-type yeast. Such an escape synthesis of the GAL enzymes was not detected if GAL4 or both GAL4 and GAL7 were carried by a single-copy plasmid. The results suggest that the escape synthesis of GAL enzymes observed in the GAL4-amplified yeast was a consequence of overproduction of the GAL4 protein. The GAL80 gene negatively regulating the GAL cluster genes was also isolated, and when amplified together with GAL4, no escape synthesis of the GAL enzymes was observed, suggesting that the balanced synthesis of two regulatory proteins was essential to maintain the repressed state of the GAL cluster genes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bassel J, Mortimer R (1971) Genetic order of the galactose structural genes in Saccharomyces cerevisiae. J Bacteriol 108:179–183

    Google Scholar 

  • Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–108

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acid Res 7:1513–1523

    Google Scholar 

  • Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW (1979) Sterile host yeasts (SHY): A eukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–24

    Google Scholar 

  • Broach JR (1979) Galactose Regulation in Saccharomyces cerevisiae. The enzymes encoded by the GAL7, 10, 1 cluster are coordinately controlled and separately translated. J Mol Biol 131:41–53

    Google Scholar 

  • Broach JR, Strathern JN, Hicks JB (1979) Transformation in Yeast: Development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133

    Google Scholar 

  • Buttin G (1963) Mecanismes régulateurs dans la biosynthése des enzymes du métabolisme du galactose chez Escherichia coli K12. J Mol Biol 7:164–182

    Google Scholar 

  • Clarke L, Carbon J (1980) Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature 287:504–509

    Google Scholar 

  • Cryer DR, Eccleshall R, Marmur J (1975) Isolation of yeast DNA. In: Prescott DM (ed) Methods in cell biology, vol 12. Academic Press, New York, p 39–44

    Google Scholar 

  • Davis RW, Thomas M, Cameron J, St. John TP, Scherer S, Padgett RA (1979) Rapid DNA isolations for enzymatic and hybridization analysis. Methods Enzymol 65:404–414

    Google Scholar 

  • Douglas HC, Hawthorne DC (1964) Enzymatic expression and genetic linkage of genes controlling galactose utilization in Saccharomyces. Genetics 49:837–844

    Google Scholar 

  • Douglas HC, Hawthorne DC (1966) Regulation of genes controlling synthesis of the galactose pathway enzymes in yeast. Genetics 54:911–916

    Google Scholar 

  • Douglas HC, Hawthorne DC (1972) Uninducible mutants in the gali locus of Saccharomyces cerevisiae. J Bacteriol 109:1139–1143

    Google Scholar 

  • Guarente L, Yocum RR, Gifford P (1982) A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. Proc Natl Acad Sci USA 79:7410–7414

    Google Scholar 

  • Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci USA 75:1929–1933

    Google Scholar 

  • Johnston SA, Hopper JE (1982) Isolation of the yeast regulatory gene GAL4 and analysis of its dosage effects on the galactose/ melibiose regulon. Proc Natl Acad Sci USA 79:6971–6975

    Google Scholar 

  • Kew OH, Douglas HC (1976) Genetic co-regulation of galactose and melibiose utilization in Saccharomyces. J Bacteriol 125:33–41

    Google Scholar 

  • Klar AJS, Halvorson HO (1974) Studies on the positive regulatory gene, GAL4, in regulation of galactose catabolic enzymes in Saccharomyces cerevisiae. Mol Gen Genet 125:203–212

    Google Scholar 

  • Laughon A, Gesteland RF (1982) Isolation and preliminary characterization of the GAL4 gene, a positive regulator of transcription in yeast. Proc Natl Acad Sci USA 79:6827–6831

    Google Scholar 

  • Matsumoto K, Toh-e A, Oshima Y (1978) Genetic control of galactokinase synthesis in Saccharomyces cerevisiae: Evidence for constitutive expression of the positive regulatory gene gal4. J Bacteriol 134:446–457

    Google Scholar 

  • Matsumoto K, Adachi Y, To-e A, Oshima Y (1980) Function of positive regulatory gene gal4 in the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae: Evidence that the GAL81 region codes for part of the gal4 protein. J Bacteriol 141:508–527

    Google Scholar 

  • Nogi Y, Fukasawa T (1980) A novel mutation that affects utilization of galactose in Saccharomyces cerevisiae. Curr Genet 2:115–120

    Google Scholar 

  • Perlman D, Hopper JE (1979) Constitutive synthesis of the GAL4 protein, a galactose pathway regulator in Saccharomyces cerevisiae. Cell 16:89–95

    Google Scholar 

  • Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1974) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251

    Google Scholar 

  • Segawa T, Fukasawa T (1980) Transcriptional units of GAL genes in Saccharomyces cerevisiae determined by ultraviolet light mapping. Curr Genet 2:223–228

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • St John TP, Davis RW (1979) Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization. Cell 16:443–452

    Google Scholar 

  • St John TP, Davis RW (1981) The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol 152:285–315

    Google Scholar 

  • St John TP, Scherer S, McDonell MW, Davis RW (1981) Deletion analysis of the Saccharomyces GAL gene cluster. J Mol Biol 152:317–334

    Google Scholar 

  • Stinchcomb DT, Mann C, Davis RW (1982) Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol 158:157–179

    Google Scholar 

  • Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) High-frequency transformation of yeast: Autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by T. Yura

On a leave absence from Nikka Whisky Co.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hashimoto, H., Kikuchi, Y., Nogi, Y. et al. Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae . Molec Gen Genet 191, 31–38 (1983). https://doi.org/10.1007/BF00330886

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330886

Keywords

Navigation