Skip to main content
Log in

Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae

II. The isolation and dosage effect of the regulatory gene GAL80

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

Summary

The galactose analogue 2-deoxygalactose was found to inhibit the growth of a mutant strain of Saccharomyces cerevisiae constitutively producing the set of galactose utilization enzymes. Based on this fact, the yeast GAL80 gene negatively regulating the expression of the genes encoding those enzymes was isolated for its ability to confer 2-deoxygalactose resistance on a strain carrying a recessive mutation in that gene. The GAL80 gene was located within a 3.0 kb fragment in the cloned DNA. When the isolated gene was incorporated into a multi-copy plasmid, the induced level of three enzymes encoded by the gene cluster GAL7-GAL10-GAL1 in the host chromosome was lowered. Such a gene dosage effect of GAL80 was further pronounced if sucrose, a sugar causing catabolite repression, was added to the growth medium. The ratio of the enzyme activity of the yeast bearing multiple copies of GAL80 to that of the yeast bearing its single copy significantly varied with the enzyme. From these results we suggest that the intracellular inducer interacts with the GAL80 product and that GAL80 molecules directly bind the GAL cluster genes with an affinity different from one gene to another.

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

  • Adams BG (1972) Induction of galactokinase in Saccharomyces cerevisiae: kinetics of induction and glucose effects. J Bacteriol 111:308–315

    Google Scholar 

  • Alper MD, Ames BN (1975) Positive selection of mutants with deletions of the gal-chl region of the Salmonella chromosome as a screening procedure for mutagens that cause deletions. J Bacteriol 121:259–266

    Google Scholar 

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

    Google Scholar 

  • Broach JR (1979) Galactose regulation in Sacchromyces 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, Stathern 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 

  • Cirillo VP (1968) Galactose transport in Saccharomyces cerevisiae: I. Nonmetabolized sugars as substrates and inducers of the galactose transport system. J Bacteriol 95:1727–1731

    Google Scholar 

  • Douglas HC, Condie F (1954) The genetic control of galactose utilization in Saccharomyces. J Bacteriol 68:662–670

    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 gal locus of Saccharomyces cervisiae. J Bacteriol 109:1139–1143

    Google Scholar 

  • Fukasawa T, Segawa T, Nogi Y (1982) Uridine diphosphate glucose-4-epimerase and galactose-1-phosphate uridylyltransferase from Saccharomyces cerevisiae. In: Wood WA (eds) Methods in enzymol. (Academic Press, New York), Vol 89, Part D, pp 584–592

    Google Scholar 

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

    Google Scholar 

  • Hashimoto H, Kikuchi Y, Nogi Y, Fukasawa T (1983) Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae: Isolation and characterization of the regulatory gene GAL4. Molec Gen Genet 191:31–38

    Google Scholar 

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

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    Google Scholar 

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

    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 

  • Matern H, Holzer H (1977) Catabolite inactivation of the galactose uptake system in yeast. J Biol Chem 252:6399–6402

    Google Scholar 

  • Matsumoto K, Adachi Y, Toh-e A, Oshima Y (1978) 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 

  • Nogi Y, Matsumoto K, Toh-e A, Oshima Y (1977) Interaction of super-repressible and dominant constitutive mutations for the synthesis of galactose pathway enzyme in Saccharomyces cerevisiae. Molec Gen Genet 152:137–144

    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 

  • 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 

  • Thirion J-P, Banville D, Noel H (1976) Galactokinase mutants of Chinese hamster somatic cells resistant to 2-deoxygalactose. Genetics 83:137–147

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communited by M. Takanami

The first article of this series is in Mol Gen Genet 191:31–38

On a leave absence from Nikka Whisky Co.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nogi, Y., Shimada, H., Matsuzaki, Y. et al. Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae . Mol Gen Genet 195, 29–34 (1984). https://doi.org/10.1007/BF00332719

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00332719

Keywords

Navigation