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. 1996 Apr;142(4):1083–1093. doi: 10.1093/genetics/142.4.1083

Rox3 and Rts1 Function in the Global Stress Response Pathway in Baker's Yeast

C C Evangelista-Jr 1, A M Rodriguez-Torres 1, M P Limbach 1, R S Zitomer 1
PMCID: PMC1207109  PMID: 8846889

Abstract

Yeast respond to a variety of stresses through a global stress response that is mediated by a number of signal transduction pathways and the cis-acting STRE DNA sequence. The CYC7 gene, encoding iso-2-cytochrome c, has been demonstrated to respond to heat shock, glucose starvation, approach-to-stationary phase, and, as we demonstrate here, to osmotic stress. This response was delayed in a the hog1-Δ1 strain implicating the Hog1 mitogen-activated protein kinase cascade, a known component of the global stress response. Deletion analysis of the CYC7 regulatory region suggested that three STRE elements were each capable of inducing the stress response. Mutations in the ROX3 gene prevented CYC7 RNA accumulation during heat shock and osmotic stress. ROX3 RNA levels were shown to be induced by stress through a novel regulatory element. A selection for high-copy suppressors of a ROX3 temperature-sensitive allele resulted in the isolation of RTS1, encoding a protein with homology to the B' regulatory subunit of protein phosphatase 2A(0). Deletion of RTS1 caused temperature and osmotic sensitivity and increased accumulation of CYC7 RNA under all conditions. Over-expression of this gene caused increased CYC7 RNA accumulation in rox3 mutants but not in wild-type cells.

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Selected References

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  1. Arndt K. T., Styles C. A., Fink G. R. A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell. 1989 Feb 24;56(4):527–537. doi: 10.1016/0092-8674(89)90576-x. [DOI] [PubMed] [Google Scholar]
  2. Craig E. A., Gambill B. D., Nelson R. J. Heat shock proteins: molecular chaperones of protein biogenesis. Microbiol Rev. 1993 Jun;57(2):402–414. doi: 10.1128/mr.57.2.402-414.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  4. Gounalaki N., Thireos G. Yap1p, a yeast transcriptional activator that mediates multidrug resistance, regulates the metabolic stress response. EMBO J. 1994 Sep 1;13(17):4036–4041. doi: 10.1002/j.1460-2075.1994.tb06720.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Healy A. M., Helser T. L., Zitomer R. S. Sequences required for transcriptional initiation of the Saccharomyces cerevisiae CYC7 genes. Mol Cell Biol. 1987 Oct;7(10):3785–3791. doi: 10.1128/mcb.7.10.3785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Healy A. M., Zolnierowicz S., Stapleton A. E., Goebl M., DePaoli-Roach A. A., Pringle J. R. CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: identification, characterization, and homology to the B subunit of mammalian type 2A protein phosphatase. Mol Cell Biol. 1991 Nov;11(11):5767–5780. doi: 10.1128/mcb.11.11.5767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hinnebusch A. G. Transcriptional and translational regulation of gene expression in the general control of amino-acid biosynthesis in Saccharomyces cerevisiae. Prog Nucleic Acid Res Mol Biol. 1990;38:195–240. doi: 10.1016/s0079-6603(08)60712-6. [DOI] [PubMed] [Google Scholar]
  8. Hodge M. R., Kim G., Singh K., Cumsky M. G. Inverse regulation of the yeast COX5 genes by oxygen and heme. Mol Cell Biol. 1989 May;9(5):1958–1964. doi: 10.1128/mcb.9.5.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Klebe R. J., Harriss J. V., Sharp Z. D., Douglas M. G. A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast. Gene. 1983 Nov;25(2-3):333–341. doi: 10.1016/0378-1119(83)90238-x. [DOI] [PubMed] [Google Scholar]
  10. Kobayashi N., McEntee K. Identification of cis and trans components of a novel heat shock stress regulatory pathway in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jan;13(1):248–256. doi: 10.1128/mcb.13.1.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Laloraya S., Gambill B. D., Craig E. A. A role for a eukaryotic GrpE-related protein, Mge1p, in protein translocation. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6481–6485. doi: 10.1073/pnas.91.14.6481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lowry C. V., Weiss J. L., Walthall D. A., Zitomer R. S. Modulator sequences mediate oxygen regulation of CYC1 and a neighboring gene in yeast. Proc Natl Acad Sci U S A. 1983 Jan;80(1):151–155. doi: 10.1073/pnas.80.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lowry C. V., Zitomer R. S. ROX1 encodes a heme-induced repression factor regulating ANB1 and CYC7 of Saccharomyces cerevisiae. Mol Cell Biol. 1988 Nov;8(11):4651–4658. doi: 10.1128/mcb.8.11.4651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lue N. F., Buchman A. R., Kornberg R. D. Activation of yeast RNA polymerase II transcription by a thymidine-rich upstream element in vitro. Proc Natl Acad Sci U S A. 1989 Jan;86(2):486–490. doi: 10.1073/pnas.86.2.486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maeda T., Tsai A. Y., Saito H. Mutations in a protein tyrosine phosphatase gene (PTP2) and a protein serine/threonine phosphatase gene (PTC1) cause a synthetic growth defect in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Sep;13(9):5408–5417. doi: 10.1128/mcb.13.9.5408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maeda T., Wurgler-Murphy S. M., Saito H. A two-component system that regulates an osmosensing MAP kinase cascade in yeast. Nature. 1994 May 19;369(6477):242–245. doi: 10.1038/369242a0. [DOI] [PubMed] [Google Scholar]
  17. Pillar T. M., Bradshaw R. E. Heat shock and stationary phase induce transcription of the Saccharomyces cerevisiae iso-2 cytochrome c gene. Curr Genet. 1991 Aug;20(3):185–188. doi: 10.1007/BF00326230. [DOI] [PubMed] [Google Scholar]
  18. Prezant T., Pfeifer K., Guarente L. Organization of the regulatory region of the yeast CYC7 gene: multiple factors are involved in regulation. Mol Cell Biol. 1987 Sep;7(9):3252–3259. doi: 10.1128/mcb.7.9.3252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ronne H., Carlberg M., Hu G. Z., Nehlin J. O. Protein phosphatase 2A in Saccharomyces cerevisiae: effects on cell growth and bud morphogenesis. Mol Cell Biol. 1991 Oct;11(10):4876–4884. doi: 10.1128/mcb.11.10.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ronne H. Glucose repression in fungi. Trends Genet. 1995 Jan;11(1):12–17. doi: 10.1016/s0168-9525(00)88980-5. [DOI] [PubMed] [Google Scholar]
  21. Rosenblum-Vos L. S., Rhodes L., Evangelista C. C., Jr, Boayke K. A., Zitomer R. S. The ROX3 gene encodes an essential nuclear protein involved in CYC7 gene expression in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Nov;11(11):5639–5647. doi: 10.1128/mcb.11.11.5639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rospert S., Junne T., Glick B. S., Schatz G. Cloning and disruption of the gene encoding yeast mitochondrial chaperonin 10, the homolog of E. coli groES. FEBS Lett. 1993 Dec 13;335(3):358–360. doi: 10.1016/0014-5793(93)80419-u. [DOI] [PubMed] [Google Scholar]
  23. Rotenberg M. O., Woolford J. L., Jr Tripartite upstream promoter element essential for expression of Saccharomyces cerevisiae ribosomal protein genes. Mol Cell Biol. 1986 Feb;6(2):674–687. doi: 10.1128/mcb.6.2.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rothstein R. J., Sherman F. Genes affecting the expression of cytochrome c in yeast: genetic mapping and genetic interactions. Genetics. 1980 Apr;94(4):871–889. doi: 10.1093/genetics/94.4.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Russell D. W., Smith M., Cox D., Williamson V. M., Young E. T. DNA sequences of two yeast promoter-up mutants. Nature. 1983 Aug 18;304(5927):652–654. doi: 10.1038/304652a0. [DOI] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schüller C., Brewster J. L., Alexander M. R., Gustin M. C., Ruis H. The HOG pathway controls osmotic regulation of transcription via the stress response element (STRE) of the Saccharomyces cerevisiae CTT1 gene. EMBO J. 1994 Sep 15;13(18):4382–4389. doi: 10.1002/j.1460-2075.1994.tb06758.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sherman F., Stewart J. W. Genetics and biosynthesis of cytochrome c. Annu Rev Genet. 1971;5:257–296. doi: 10.1146/annurev.ge.05.120171.001353. [DOI] [PubMed] [Google Scholar]
  29. Shu Y., Hallberg R. L. SCS1, a multicopy suppressor of hsp60-ts mutant alleles, does not encode a mitochondrially targeted protein. Mol Cell Biol. 1995 Oct;15(10):5618–5626. doi: 10.1128/mcb.15.10.5618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sneddon A. A., Cohen P. T., Stark M. J. Saccharomyces cerevisiae protein phosphatase 2A performs an essential cellular function and is encoded by two genes. EMBO J. 1990 Dec;9(13):4339–4346. doi: 10.1002/j.1460-2075.1990.tb07883.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sutton A., Immanuel D., Arndt K. T. The SIT4 protein phosphatase functions in late G1 for progression into S phase. Mol Cell Biol. 1991 Apr;11(4):2133–2148. doi: 10.1128/mcb.11.4.2133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Waterland R. A., Basu A., Chance B., Poyton R. O. The isoforms of yeast cytochrome c oxidase subunit V alter the in vivo kinetic properties of the holoenzyme. J Biol Chem. 1991 Mar 5;266(7):4180–4186. [PubMed] [Google Scholar]
  33. Werner-Washburne M., Braun E., Johnston G. C., Singer R. A. Stationary phase in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1993 Jun;57(2):383–401. doi: 10.1128/mr.57.2.383-401.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wright C. F., Zitomer R. S. A positive regulatory site and a negative regulatory site control the expression of the Saccharomyces cerevisiae CYC7 gene. Mol Cell Biol. 1984 Oct;4(10):2023–2030. doi: 10.1128/mcb.4.10.2023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zitomer R. S., Sellers J. W., McCarter D. W., Hastings G. A., Wick P., Lowry C. V. Elements involved in oxygen regulation of the Saccharomyces cerevisiae CYC7 gene. Mol Cell Biol. 1987 Jun;7(6):2212–2220. doi: 10.1128/mcb.7.6.2212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zolnierowicz S., Csortos C., Bondor J., Verin A., Mumby M. C., DePaoli-Roach A. A. Diversity in the regulatory B-subunits of protein phosphatase 2A: identification of a novel isoform highly expressed in brain. Biochemistry. 1994 Oct 4;33(39):11858–11867. doi: 10.1021/bi00205a023. [DOI] [PubMed] [Google Scholar]
  37. van Zyl W., Huang W., Sneddon A. A., Stark M., Camier S., Werner M., Marck C., Sentenac A., Broach J. R. Inactivation of the protein phosphatase 2A regulatory subunit A results in morphological and transcriptional defects in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Nov;12(11):4946–4959. doi: 10.1128/mcb.12.11.4946. [DOI] [PMC free article] [PubMed] [Google Scholar]

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