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Transcription of multiple copies of the yeast GAL7 gene is limited by specific factors in addition to GAL4

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Summary

High levels of the GAL7 gene in the yeast cell appear to titrate regulatory factors and to impair transcription of related sequences. To investigate the role that the GAL regulatory factors GAL4 and GAL80 have in this process we have compared the accumulation of mRNA transcribed from single-copy (plasmid-borne GAL7 and chromosomal GAL10) and high-copy (plasmid-borne GAL7) genes in several GAL regulatory mutants. Our results show that functional GAL4 gene product is required for induction of transcription from the single- and high-copy genes. In a strain containing the GAL4 gene fused to the high expression ADH1 promoter, glucose can replace galactose to induce high levels of transcription of GAL7 and GAL10 genes, although the kinetics of accumulation induced by the two sugars are distinctly different. In the presence of high levels of GAL4, maximum accumulation of mRNA from single and high copy genes is elevated two-fold; disruption of the gal80 gene in combination with high levels of GAL4 results in a further two-fold increase in transcription. In this genetic background, galactose-induced transcription of the high copy GAL7 gene results in a greater than 50-fold increase in the levels of GAL7 mRNA, representing 30%–50% of the total cellular mRNA. Our results are consistent with a cooperative effect of saturation of multiple GAL4 DNA binding sites and with a limiting factor, in addition to GAL4, that is required for transcription of the GAL genes.

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References

  • Ammerer G (1983) Expression of genes in yeast using the ADC1 promoter. Methods Enzymol 101:192–201

    Google Scholar 

  • Bach ML, Lacroute F, Botstein D (1979) Evidence for transcriptional regulation of orotidine-5′-phosphate decarboxylase in yeast by hybridization of mRNA to the yeast structural gene cloned in Escherichia coli. Proc Natl Acad Sci USA 76:386–390

    Google Scholar 

  • Baker SM, Okkema PG, Jaehning JA (1984) Expression of the Saccharomyces cerevisiae GAL7 gene on autonomously replicating plasmids. Mol Cell Biol 4:2062–2071

    Google Scholar 

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

    Google Scholar 

  • Bram R, Kornberg R (1985) Specific protein binding to far upstream activating sequences in polymerase II promoters. Proc Natl Acad Sci USA 82:43–47

    Google Scholar 

  • Broach JR (1983) Construction of high copy yeast vectors using 2 μm circle sequences. Methods Enzymol 100:293–308

    Google Scholar 

  • Carlson M, Botstein D (1982) Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase. Cell 28:145–154

    Google Scholar 

  • Denis CL, Ferguson J, Young ET (1983) mRNA level for the fermentative alcohol dehydrogenase of Saccharomyces cerevisiae decreases upon growth on a nonfermentable carbon source. J Biol Chem 258:1165–1171

    Google Scholar 

  • Dobson MJ, Futcher AB, Cox BS (1980) Loss of 2μm DNA from Saccharomyces cerevisiae transformed with the chimeric plasmid pJDB219. Curr Genet 2:201–215

    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 

  • Erhart E, Hollenberg CP (1983) The presence of a defective LEU2 gene on 2 μm recombinant plasmids of Saccharomyces cerevisiae is responsible for curing and high copy number. J Bacteriol 156:625–635

    Google Scholar 

  • Falco C, Li YY, Broach JR, Botstein D (1982) Genetic properties of chromosomally integrated 2 μ plasmid DNA in yeast. Cell 29:573–584

    Google Scholar 

  • Frischauf AM, Gasoff H, Lehrach H (1980) A subcloning strategy for DNA sequence analysis. Nucleic Acids Res 8:5541–5549

    Google Scholar 

  • Giniger E, Varnum SM, Ptashne M (1985) Specific DNA binding of GAL4, a positive regulatory protein in yeast. Cell 40:767–774

    Google Scholar 

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

    Google Scholar 

  • Hopper JE, Broach J, Rowe L (1978) Regulation of the galactose pathway in Saccharomyces cerevisiae: induction of uridyl transferase mRNA and dependency of GAL4 gene function. Proc Natl Acad Sci USA 75:2878–2882

    Google Scholar 

  • Jensen R, Sprague GF, Herskowitz I (1983) Regulation of yeast mating type conversion: evidence for feedback control of HO gene expression by the mating type locus. Proc Natl Acad Sci USA 80:3035–3039

    Google Scholar 

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

    Google Scholar 

  • Johnston SA, Zavortink MJ, Debouck C, Rosenberg M, Hopper JE (1986) Functional domains of the yeast regulatory protein, GAL4. Proc Natl Acad Sci USA 83:6553–6557

    Google Scholar 

  • Keegan L, Gill G, Ptashne M (1986) Separation of DNA binding from the transcription-activating function of a eukaryotic regulatory protein. Science 231:699–704

    Google Scholar 

  • Laughon A, Driscoll R, Wills N, Gesteland R (1984) Identification of two proteins encoded by the Saccharomyces cerevisiae GAL4 gene. Mol Cell Biol 4:268–275

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Press, Cold Spring Harbor, NY

    Google Scholar 

  • Matsumoto K, Toh-e A, Oshima Y (1981) Isolation and characterization of dominant mutants resistant to carbon catabolite repression of galactokinase synthesis in Saccharomyces cerevisiae. Mol Cell Biol 1:83–93

    Google Scholar 

  • Matsumoto K, Yoshimatsu T, Oshima Y (1983) Recessive mutations conferring resistance to carbon catabolite repression of galactokinase synthesis in Saccharomyces cerevisiae. J Bacteriol 153:1405–1414

    Google Scholar 

  • Ng R, Abelson J (1980) Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 77:3912–3916

    Google Scholar 

  • Nogi Y, Fukasawa T (1984) Nucleotide sequence of the yeast regulatory gene GAL80. Nucleic Acids Res 12:9287–9298

    Google Scholar 

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

    Google Scholar 

  • Oshima Y (1982) Regulatory circuts for gene expression: the metabolism of galactose and phosphate. In: Strathern JA, Jones EW, Broach JR (eds) The Molecular Biology of the Yeast Saccharomyces, Vol. II. Cold Spring Harbor Press, Cold Spring Harbor, NY, pp 159–180

    Google Scholar 

  • Platt T (1984) Toxicity of 2-deoxygalactose to Saccharomyces cerevisiae cells constitutively synthesizing galactose metabolizing enzymes. Mol Cell Biol 4:994–996

    Google Scholar 

  • Sherman FG, Fink G, Lawrence CW (1979) Methods in Yeast Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    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 (1981) The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol 152:285–315

    Google Scholar 

  • Struhl K (1982) The yeast HIS3 promoter contains at least two distinct elements. Proc Natl Acad Sci USA 79:7385–7389

    Google Scholar 

  • Torchia TE, Hamilton RW, Cano CL, Hopper JE (1984) Disruption of regulatory gene GAL80 in Saccharomyces cerevisiae: effects of carbon-controlled regulation of the galactose/melibiose pathway genes. Mol Cell Biol 4:1521–1527

    Google Scholar 

  • West RW Jr, Yocum R, Ptashne M (1984) Saccharomyces cerevisiae GAL1-GAL10 divergent promoter region: location and function of the upstream activating sequence UASG. Mol Cell Biol 4:2467–2478

    Google Scholar 

  • Yocum RR, Johnston M (1984) Molecular cloning of the GAL80 gene from Saccharomyces cerevisiae and characterization of a gal80 deletion. Gene 32:75–82

    Google Scholar 

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Communicated by C.P. Hollenberg

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Baker, S.M., Johnston, S.A., Hopper, J.E. et al. Transcription of multiple copies of the yeast GAL7 gene is limited by specific factors in addition to GAL4 . Mole Gen Genet 208, 127–134 (1987). https://doi.org/10.1007/BF00330433

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  • DOI: https://doi.org/10.1007/BF00330433

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