Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Jun;87(11):4098–4102. doi: 10.1073/pnas.87.11.4098

cAMP-dependent phosphorylation and inactivation of yeast transcription factor ADR1 does not affect DNA binding.

W E Taylor 1, E T Young 1
PMCID: PMC54054  PMID: 2161531

Abstract

Transcription factor ADR1 increases the level of ADH2 gene expression 200-fold by binding to a palindromic upstream activation sequence (UAS1) in the glucose-repressible ADH2 promoter in Saccharomyces cerevisiae. cAMP-dependent protein kinase (cAPK) phosphorylates ADR1 in vitro and a yeast strain with elevated cAPK activity inhibits the ability of ADR1 to activate ADH2 transcription in vivo [Cherry, J. R., Johnson, T. R., Dollard, C., Schuster, J. R. & Denis, C. L. (1988) Cell 56, 409-419]. Intact ADR1 protein was detected at comparable levels in extracts made from repressed or derepressed yeast cells, indicating that glucose repression is not due to absence of ADR1. ADR1 in extracts made from glucose-repressed and -derepressed cells bound UAS1 DNA with similar affinities despite having greatly different abilities to activate ADH2 gene expression in vivo. A mutant form of ADR1 encoded by ADR1-5c, which has an altered consensus sequence for phosphorylation by cAPK conferred constitutive expression on ADH2 but bound DNA to the same extent as wild-type ADR1 protein. Similarly, normal DNA binding was seen for ADR1 produced in mutants with altered levels of cAPK activity. Because inactivation of ADR1 by phosphorylation has no detectable effect on either DNA binding or ADR1 levels, ADR1 probably binds to UAS1 constitutively and phosphorylation prevents it from promoting transcription.

Full text

PDF
4098

Images in this article

4099Image
on p.4099
4099Image
on p.4099
4100Image
on p.4100
4100Image
on p.4100
4101Image
on p.4101
4101Image
on p.4101
4101Image
on p.4101

Selected References

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

  1. Ammerer G. Expression of genes in yeast using the ADCI promoter. Methods Enzymol. 1983;101:192–201. doi: 10.1016/0076-6879(83)01014-9. [DOI] [PubMed] [Google Scholar]
  2. Bissinger P. H., Wieser R., Hamilton B., Ruis H. Control of Saccharomyces cerevisiae catalase T gene (CTT1) expression by nutrient supply via the RAS-cyclic AMP pathway. Mol Cell Biol. 1989 Mar;9(3):1309–1315. doi: 10.1128/mcb.9.3.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blumberg H., Eisen A., Sledziewski A., Bader D., Young E. T. Two zinc fingers of a yeast regulatory protein shown by genetic evidence to be essential for its function. 1987 Jul 30-Aug 5Nature. 328(6129):443–445. doi: 10.1038/328443a0. [DOI] [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. Cannon J. F., Tatchell K. Characterization of Saccharomyces cerevisiae genes encoding subunits of cyclic AMP-dependent protein kinase. Mol Cell Biol. 1987 Aug;7(8):2653–2663. doi: 10.1128/mcb.7.8.2653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Celenza J. L., Carlson M. A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science. 1986 Sep 12;233(4769):1175–1180. doi: 10.1126/science.3526554. [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. 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]
  9. 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]
  10. 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]
  11. 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]
  12. 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]
  13. 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]
  14. Eisen A., Taylor W. E., Blumberg H., Young E. T. The yeast regulatory protein ADR1 binds in a zinc-dependent manner to the upstream activating sequence of ADH2. Mol Cell Biol. 1988 Oct;8(10):4552–4556. doi: 10.1128/mcb.8.10.4552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Irani M., Taylor W. E., Young E. T. Transcription of the ADH2 gene in Saccharomyces cerevisiae is limited by positive factors that bind competitively to its intact promoter region on multicopy plasmids. Mol Cell Biol. 1987 Mar;7(3):1233–1241. doi: 10.1128/mcb.7.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kemp B. E., Graves D. J., Benjamini E., Krebs E. G. Role of multiple basic residues in determining the substrate specificity of cyclic AMP-dependent protein kinase. J Biol Chem. 1977 Jul 25;252(14):4888–4894. [PubMed] [Google Scholar]
  19. Levitzki A. Transmembrane signalling to adenylate cyclase in mammalian cells and in Saccharomyces cerevisiae. Trends Biochem Sci. 1988 Aug;13(8):298–301. doi: 10.1016/0968-0004(88)90122-3. [DOI] [PubMed] [Google Scholar]
  20. Matsumoto K., Uno I., Oshima Y., Ishikawa T. Isolation and characterization of yeast mutants deficient in adenylate cyclase and cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2355–2359. doi: 10.1073/pnas.79.7.2355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Montminy M. R., Bilezikjian L. M. Binding of a nuclear protein to the cyclic-AMP response element of the somatostatin gene. Nature. 1987 Jul 9;328(6126):175–178. doi: 10.1038/328175a0. [DOI] [PubMed] [Google Scholar]
  22. Párraga G., Horvath S. J., Eisen A., Taylor W. E., Hood L., Young E. T., Klevit R. E. Zinc-dependent structure of a single-finger domain of yeast ADR1. Science. 1988 Sep 16;241(4872):1489–1492. doi: 10.1126/science.3047872. [DOI] [PubMed] [Google Scholar]
  23. Rittenhouse J., Moberly L., Marcus F. Phosphorylation in vivo of yeast (Saccharomyces cerevisiae) fructose-1,6-bisphosphatase at the cyclic AMP-dependent site. J Biol Chem. 1987 Jul 25;262(21):10114–10119. [PubMed] [Google Scholar]
  24. Scott J. D., Fischer E. H., Takio K., Demaille J. G., Krebs E. G. Amino acid sequence of the heat-stable inhibitor of the cAMP-dependent protein kinase from rabbit skeletal muscle. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5732–5736. doi: 10.1073/pnas.82.17.5732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shuster J., Yu J., Cox D., Chan R. V., Smith M., Young E. ADR1-mediated regulation of ADH2 requires an inverted repeat sequence. Mol Cell Biol. 1986 Jun;6(6):1894–1902. doi: 10.1128/mcb.6.6.1894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Tanaka K., Matsumoto K., Toh-e A. Dual regulation of the expression of the polyubiquitin gene by cyclic AMP and heat shock in yeast. EMBO J. 1988 Feb;7(2):495–502. doi: 10.1002/j.1460-2075.1988.tb02837.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thukral S. K., Tavianini M. A., Blumberg H., Young E. T. Localization of a minimal binding domain and activation regions in yeast regulatory protein ADR1. Mol Cell Biol. 1989 Jun;9(6):2360–2369. doi: 10.1128/mcb.9.6.2360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Toda T., Cameron S., Sass P., Zoller M., Scott J. D., McMullen B., Hurwitz M., Krebs E. G., Wigler M. Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Apr;7(4):1371–1377. doi: 10.1128/mcb.7.4.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yamamoto K. K., Gonzalez G. A., Biggs W. H., 3rd, Montminy M. R. Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature. 1988 Aug 11;334(6182):494–498. doi: 10.1038/334494a0. [DOI] [PubMed] [Google Scholar]
  31. 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 Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES