Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1992 Apr;12(4):1663–1673. doi: 10.1128/mcb.12.4.1663

Glucose repression of the yeast ADH2 gene occurs through multiple mechanisms, including control of the protein synthesis of its transcriptional activator, ADR1.

R C Vallari 1, W J Cook 1, D C Audino 1, M J Morgan 1, D E Jensen 1, A P Laudano 1, C L Denis 1
PMCID: PMC369609  PMID: 1549119

Abstract

The rate of ADH2 transcription increases dramatically when Saccharomyces cerevisiae cells are shifted from glucose to ethanol growth conditions. Since ADH2 expression under glucose growth conditions is strictly dependent on the dosage of the transcriptional activator ADR1, we investigated the possibility that regulation of the rate of ADR1 protein synthesis plays a role in controlling ADR1 activation of ADH2 transcription. We found that the rate of ADR1 protein synthesis increased 10- to 16-fold within 40 to 60 min after glucose depletion, coterminous with initiation of ADH2 transcription. Changes in ADR1 mRNA levels contributed only a twofold effect on ADR1 protein synthetic differences. The 510-nt untranslated ADR1 mRNA leader sequence was found to have no involvement in regulating the rate of ADR1 protein synthesis. In contrast, sequences internal to ADR1 coding region were determined to be necessary for controlling ADR1 translation. The ADR1c mutations which enhance ADR1 activity under glucose growth conditions did not affect ADR1 protein translation. ADR1 was also shown to be multiply phosphorylated in vivo under both ethanol and glucose growth conditions. Our results indicate that derepression of ADH2 occurs through multiple mechanisms involving the ADR1 regulatory protein.

Full text

PDF
1663

Images in this article

1666Image
on p.1666
1666Image
on p.1666
1667Image
on p.1667
1667Image
on p.1667
1667Image
on p.1667
1669Image
on p.1669
1669Image
on p.1669
1670Image
on p.1670
1670Image
on p.1670
1671Image
on p.1671

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barber J. R., Verma I. M. Modification of fos proteins: phosphorylation of c-fos, but not v-fos, is stimulated by 12-tetradecanoyl-phorbol-13-acetate and serum. Mol Cell Biol. 1987 Jun;7(6):2201–2211. doi: 10.1128/mcb.7.6.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bemis L. T., Denis C. L. Characterization of the adr1-1 nonsense mutation identifies the translational start of the yeast transcriptional activator ADR1. Yeast. 1989 Jul-Aug;5(4):291–298. doi: 10.1002/yea.320050409. [DOI] [PubMed] [Google Scholar]
  3. Bemis L. T., Denis C. L. Identification of functional regions in the yeast transcriptional activator ADR1. Mol Cell Biol. 1988 May;8(5):2125–2131. doi: 10.1128/mcb.8.5.2125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blumberg H., Hartshorne T. A., Young E. T. Regulation of expression and activity of the yeast transcription factor ADR1. Mol Cell Biol. 1988 May;8(5):1868–1876. doi: 10.1128/mcb.8.5.1868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bostian K. A., Lemire J. M., Cannon L. E., Halvorson H. O. In vitro synthesis of repressible yeast acid phosphatase: identification of multiple mRNAs and products. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4504–4508. doi: 10.1073/pnas.77.8.4504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cherry J. R., Denis C. L. Overexpression of the yeast transcriptional activator ADR1 induces mutation of the mitochondrial genome. Curr Genet. 1989 May;15(5):311–317. doi: 10.1007/BF00419910. [DOI] [PubMed] [Google Scholar]
  7. Cherry J. R., Johnson T. R., Dollard C., Shuster J. R., Denis C. L. Cyclic AMP-dependent protein kinase phosphorylates and inactivates the yeast transcriptional activator ADR1. Cell. 1989 Feb 10;56(3):409–419. doi: 10.1016/0092-8674(89)90244-4. [DOI] [PubMed] [Google Scholar]
  8. Ciriacy M. Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. II. Two loci controlling synthesis of the glucose-repressible ADH II. Mol Gen Genet. 1975;138(2):157–164. doi: 10.1007/BF02428119. [DOI] [PubMed] [Google Scholar]
  9. Ciriacy M. Isolation and characterization of further cis- and trans-acting regulatory elements involved in the synthesis of glucose-repressible alcohol dehydrogenase (ADHII) in Saccharomyces cerevisiae. Mol Gen Genet. 1979 Nov;176(3):427–431. doi: 10.1007/BF00333107. [DOI] [PubMed] [Google Scholar]
  10. Ciriacy M. Isolation and characterization of yeast mutants defective in intermediary carbon metabolism and in carbon catabolite derepression. Mol Gen Genet. 1977 Jul 20;154(2):213–220. doi: 10.1007/BF00330840. [DOI] [PubMed] [Google Scholar]
  11. Denis C. L., Audino D. C. The CCR1 (SNF1) and SCH9 protein kinases act independently of cAMP-dependent protein kinase and the transcriptional activator ADR1 in controlling yeast ADH2 expression. Mol Gen Genet. 1991 Oct;229(3):395–399. doi: 10.1007/BF00267461. [DOI] [PubMed] [Google Scholar]
  12. Denis C. L., Ciriacy M., Young E. T. A positive regulatory gene is required for accumulation of the functional messenger RNA for the glucose-repressible alcohol dehydrogenase from Saccharomyces cerevisiae. J Mol Biol. 1981 Jun 5;148(4):355–368. doi: 10.1016/0022-2836(81)90181-9. [DOI] [PubMed] [Google Scholar]
  13. Denis C. L., Ferguson J., Young E. T. mRNA levels for the fermentative alcohol dehydrogenase of Saccharomyces cerevisiae decrease upon growth on a nonfermentable carbon source. J Biol Chem. 1983 Jan 25;258(2):1165–1171. [PubMed] [Google Scholar]
  14. Denis C. L., Fontaine S. C., Chase D., Kemp B. E., Bemis L. T. ADR1c mutations enhance the ability of ADR1 to activate transcription by a mechanism that is independent of effects on cyclic AMP-dependent protein kinase phosphorylation of Ser-230. Mol Cell Biol. 1992 Apr;12(4):1507–1514. doi: 10.1128/mcb.12.4.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Denis C. L., Gallo C. Constitutive RNA synthesis for the yeast activator ADR1 and identification of the ADR1-5c mutation: implications in posttranslational control of ADR1. Mol Cell Biol. 1986 Nov;6(11):4026–4030. doi: 10.1128/mcb.6.11.4026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Denis C. L. Identification of new genes involved in the regulation of yeast alcohol dehydrogenase II. Genetics. 1984 Dec;108(4):833–844. doi: 10.1093/genetics/108.4.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Denis C. L., Malvar T. The CCR4 gene from Saccharomyces cerevisiae is required for both nonfermentative and spt-mediated gene expression. Genetics. 1990 Feb;124(2):283–291. doi: 10.1093/genetics/124.2.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Denis C. L. The effects of ADR1 and CCR1 gene dosage on the regulation of the glucose-repressible alcohol dehydrogenase from Saccharomyces cerevisiae. Mol Gen Genet. 1987 Jun;208(1-2):101–106. doi: 10.1007/BF00330429. [DOI] [PubMed] [Google Scholar]
  19. Denis C. L., Young E. T. Isolation and characterization of the positive regulatory gene ADR1 from Saccharomyces cerevisiae. Mol Cell Biol. 1983 Mar;3(3):360–370. doi: 10.1128/mcb.3.3.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Flick J. S., Johnston M. Two systems of glucose repression of the GAL1 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Sep;10(9):4757–4769. doi: 10.1128/mcb.10.9.4757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gill G., Ptashne M. Negative effect of the transcriptional activator GAL4. Nature. 1988 Aug 25;334(6184):721–724. doi: 10.1038/334721a0. [DOI] [PubMed] [Google Scholar]
  22. Hartshorne T. A., Blumberg H., Young E. T. Sequence homology of the yeast regulatory protein ADR1 with Xenopus transcription factor TFIIIA. Nature. 1986 Mar 20;320(6059):283–287. doi: 10.1038/320283a0. [DOI] [PubMed] [Google Scholar]
  23. Kief D. R., Warner J. R. Coordinate control of syntheses of ribosomal ribonucleic acid and ribosomal proteins during nutritional shift-up in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Nov;1(11):1007–1015. doi: 10.1128/mcb.1.11.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Laudano A. P., Buchanan J. M. Phosphorylation of tyrosine in the carboxyl-terminal tryptic peptide of pp60c-src. Proc Natl Acad Sci U S A. 1986 Feb;83(4):892–896. doi: 10.1073/pnas.83.4.892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liu F. T., Zinnecker M., Hamaoka T., Katz D. H. New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates. Biochemistry. 1979 Feb 20;18(4):690–693. doi: 10.1021/bi00571a022. [DOI] [PubMed] [Google Scholar]
  26. Miller P. F., Hinnebusch A. G. Sequences that surround the stop codons of upstream open reading frames in GCN4 mRNA determine their distinct functions in translational control. Genes Dev. 1989 Aug;3(8):1217–1225. doi: 10.1101/gad.3.8.1217. [DOI] [PubMed] [Google Scholar]
  27. Reid G. A. Pulse labeling of yeast cells and spheroplasts. Methods Enzymol. 1983;97:324–329. doi: 10.1016/0076-6879(83)97144-6. [DOI] [PubMed] [Google Scholar]
  28. Simon M., Adam G., Rapatz W., Spevak W., Ruis H. The Saccharomyces cerevisiae ADR1 gene is a positive regulator of transcription of genes encoding peroxisomal proteins. Mol Cell Biol. 1991 Feb;11(2):699–704. doi: 10.1128/mcb.11.2.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Taguchi A. K., Young E. T. The identification and characterization of ADR6, a gene required for sporulation and for expression of the alcohol dehydrogenase II isozyme from Saccharomyces cerevisiae. Genetics. 1987 Aug;116(4):523–530. doi: 10.1093/genetics/116.4.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Taylor W. E., Young E. T. cAMP-dependent phosphorylation and inactivation of yeast transcription factor ADR1 does not affect DNA binding. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4098–4102. doi: 10.1073/pnas.87.11.4098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Walter G., Scheidtmann K. H., Carbone A., Laudano A. P., Doolittle R. F. Antibodies specific for the carboxy- and amino-terminal regions of simian virus 40 large tumor antigen. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5197–5200. doi: 10.1073/pnas.77.9.5197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Williamson V. M., Cox D., Young E. T., Russell D. W., Smith M. Characterization of transposable element-associated mutations that alter yeast alcohol dehydrogenase II expression. Mol Cell Biol. 1983 Jan;3(1):20–31. doi: 10.1128/mcb.3.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wills C. Regulation of sugar and ethanol metabolism in Saccharomyces cerevisiae. Crit Rev Biochem Mol Biol. 1990;25(4):245–280. doi: 10.3109/10409239009090611. [DOI] [PubMed] [Google Scholar]
  34. Yu J., Donoviel M. S., Young E. T. Adjacent upstream activation sequence elements synergistically regulate transcription of ADH2 in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Jan;9(1):34–42. doi: 10.1128/mcb.9.1.34. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES