Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1990 Aug;9(8):2523–2528. doi: 10.1002/j.1460-2075.1990.tb07432.x

Role of trans-activating proteins in the generation of active chromatin at the PHO5 promoter in S. cerevisiae.

K D Fascher 1, J Schmitz 1, W Hörz 1
PMCID: PMC552282  PMID: 2196175

Abstract

Induction of the PHO5 gene in Saccharomyces cerevisiae by phosphate starvation was previously shown to be accompanied by the removal of four positioned nucleosomes from the promoter. We have now investigated the role of two trans-activating proteins, encoded by PHO2 and PHO4, which bind to the PHO5 promoter. Both proteins are absolutely required for the chromatin transition to occur as shown by analysis of null mutants of the two genes. Transformation of these mutant strains with plasmids containing the respective genes restores the wild type chromatin response. Increasing the gene dosage of PHO2 and of PHO4 makes it possible to differentiate functionally between the two proteins. From over-expressing PHO4 in a wild type and also in a pho2 null mutant strain and complementary experiments with PHO2, it is concluded that the PHO4 protein is the primary trigger for the chromatin transition, consistent with one of its two binding sites being located between positioned nucleosomes in repressed chromatin and thereby accessible. PHO2, the binding site of which is located within a nucleosome under conditions of PHO5 repression, contributes to the chromatin transition either by destabilizing histone-DNA interactions or by under-going interactions with PHO4.

Full text

PDF
2523

Images in this article

Selected References

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

  1. Almer A., Hörz W. Nuclease hypersensitive regions with adjacent positioned nucleosomes mark the gene boundaries of the PHO5/PHO3 locus in yeast. EMBO J. 1986 Oct;5(10):2681–2687. doi: 10.1002/j.1460-2075.1986.tb04551.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Almer A., Rudolph H., Hinnen A., Hörz W. Removal of positioned nucleosomes from the yeast PHO5 promoter upon PHO5 induction releases additional upstream activating DNA elements. EMBO J. 1986 Oct;5(10):2689–2696. doi: 10.1002/j.1460-2075.1986.tb04552.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arndt K. T., Styles C., Fink G. R. Multiple global regulators control HIS4 transcription in yeast. Science. 1987 Aug 21;237(4817):874–880. doi: 10.1126/science.3303332. [DOI] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Braus G., Mösch H. U., Vogel K., Hinnen A., Hütter R. Interpathway regulation of the TRP4 gene of yeast. EMBO J. 1989 Mar;8(3):939–945. doi: 10.1002/j.1460-2075.1989.tb03455.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brendel V., Karlin S. Association of charge clusters with functional domains of cellular transcription factors. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5698–5702. doi: 10.1073/pnas.86.15.5698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Broach J. R., Strathern J. N., Hicks J. B. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene. 1979 Dec;8(1):121–133. doi: 10.1016/0378-1119(79)90012-x. [DOI] [PubMed] [Google Scholar]
  8. Bürglin T. R. The yeast regulatory gene PHO2 encodes a homeo box. Cell. 1988 May 6;53(3):339–340. doi: 10.1016/0092-8674(88)90153-5. [DOI] [PubMed] [Google Scholar]
  9. Eissenberg J. C., Cartwright I. L., Thomas G. H., Elgin S. C. Selected topics in chromatin structure. Annu Rev Genet. 1985;19:485–536. doi: 10.1146/annurev.ge.19.120185.002413. [DOI] [PubMed] [Google Scholar]
  10. Elgin S. C. The formation and function of DNase I hypersensitive sites in the process of gene activation. J Biol Chem. 1988 Dec 25;263(36):19259–19262. [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Gross D. S., Garrard W. T. Nuclease hypersensitive sites in chromatin. Annu Rev Biochem. 1988;57:159–197. doi: 10.1146/annurev.bi.57.070188.001111. [DOI] [PubMed] [Google Scholar]
  13. Haguenauer-Tsapis R., Hinnen A. A deletion that includes the signal peptidase cleavage site impairs processing, glycosylation, and secretion of cell surface yeast acid phosphatase. Mol Cell Biol. 1984 Dec;4(12):2668–2675. doi: 10.1128/mcb.4.12.2668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Han M., Kim U. J., Kayne P., Grunstein M. Depletion of histone H4 and nucleosomes activates the PHO5 gene in Saccharomyces cerevisiae. EMBO J. 1988 Jul;7(7):2221–2228. doi: 10.1002/j.1460-2075.1988.tb03061.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Kayne P. S., Kim U. J., Han M., Mullen J. R., Yoshizaki F., Grunstein M. Extremely conserved histone H4 N terminus is dispensable for growth but essential for repressing the silent mating loci in yeast. Cell. 1988 Oct 7;55(1):27–39. doi: 10.1016/0092-8674(88)90006-2. [DOI] [PubMed] [Google Scholar]
  17. Koren R., LeVitre J., Bostian K. A. Isolation of the positive-acting regulatory gene PHO4 from Saccharomyces cerevisiae. Gene. 1986;41(2-3):271–280. doi: 10.1016/0378-1119(86)90107-1. [DOI] [PubMed] [Google Scholar]
  18. Legrain M., De Wilde M., Hilger F. Isolation, physical characterization and expression analysis of the Saccharomyces cerevisiae positive regulatory gene PHO4. Nucleic Acids Res. 1986 Apr 11;14(7):3059–3073. doi: 10.1093/nar/14.7.3059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lorch Y., LaPointe J. W., Kornberg R. D. Nucleosomes inhibit the initiation of transcription but allow chain elongation with the displacement of histones. Cell. 1987 Apr 24;49(2):203–210. doi: 10.1016/0092-8674(87)90561-7. [DOI] [PubMed] [Google Scholar]
  20. Meyhack B., Bajwa W., Rudolph H., Hinnen A. Two yeast acid phosphatase structural genes are the result of a tandem duplication and show different degrees of homology in their promoter and coding sequences. EMBO J. 1982;1(6):675–680. doi: 10.1002/j.1460-2075.1982.tb01229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Panzeri L., Philippsen P. Centromeric DNA from chromosome VI in Saccharomyces cerevisiae strains. EMBO J. 1982;1(12):1605–1611. doi: 10.1002/j.1460-2075.1982.tb01362.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pederson D. S., Thoma F., Simpson R. T. Core particle, fiber, and transcriptionally active chromatin structure. Annu Rev Cell Biol. 1986;2:117–147. doi: 10.1146/annurev.cb.02.110186.001001. [DOI] [PubMed] [Google Scholar]
  23. Rudolph H., Hinnen A. The yeast PHO5 promoter: phosphate-control elements and sequences mediating mRNA start-site selection. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1340–1344. doi: 10.1073/pnas.84.5.1340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sengstag C., Hinnen A. A 28-bp segment of the Saccharomyces cerevisiae PHO5 upstream activator sequence confers phosphate control to the CYC1-lacZ gene fusion. Gene. 1988 Jul 30;67(2):223–228. doi: 10.1016/0378-1119(88)90399-x. [DOI] [PubMed] [Google Scholar]
  25. Sengstag C., Hinnen A. The sequence of the Saccharomyces cerevisiae gene PHO2 codes for a regulatory protein with unusual aminoacid composition. Nucleic Acids Res. 1987 Jan 12;15(1):233–246. doi: 10.1093/nar/15.1.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Struhl K. Molecular mechanisms of transcriptional regulation in yeast. Annu Rev Biochem. 1989;58:1051–1077. doi: 10.1146/annurev.bi.58.070189.005155. [DOI] [PubMed] [Google Scholar]
  27. Tschumper G., Carbon J. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene. Gene. 1980 Jul;10(2):157–166. doi: 10.1016/0378-1119(80)90133-x. [DOI] [PubMed] [Google Scholar]
  28. Vogel K., Hörz W., Hinnen A. The two positively acting regulatory proteins PHO2 and PHO4 physically interact with PHO5 upstream activation regions. Mol Cell Biol. 1989 May;9(5):2050–2057. doi: 10.1128/mcb.9.5.2050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Workman J. L., Roeder R. G. Binding of transcription factor TFIID to the major late promoter during in vitro nucleosome assembly potentiates subsequent initiation by RNA polymerase II. Cell. 1987 Nov 20;51(4):613–622. doi: 10.1016/0092-8674(87)90130-9. [DOI] [PubMed] [Google Scholar]
  30. Yaniv M., Cereghini S. Structure of transcriptionally active chromatin. CRC Crit Rev Biochem. 1986;21(1):1–26. doi: 10.3109/10409238609113607. [DOI] [PubMed] [Google Scholar]
  31. Yoshida K., Kuromitsu Z., Ogawa N., Oshima Y. Mode of expression of the positive regulatory genes PHO2 and PHO4 of the phosphatase regulon in Saccharomyces cerevisiae. Mol Gen Genet. 1989 May;217(1):31–39. doi: 10.1007/BF00330939. [DOI] [PubMed] [Google Scholar]
  32. Yoshida K., Ogawa N., Oshima Y. Function of the PHO regulatory genes for repressible acid phosphatase synthesis in Saccharomyces cerevisiae. Mol Gen Genet. 1989 May;217(1):40–46. doi: 10.1007/BF00330940. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES