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. 1998 Sep;150(1):31–42. doi: 10.1093/genetics/150.1.31

Identification of a calcineurin-independent pathway required for sodium ion stress response in Saccharomyces cerevisiae.

R W Ganster 1, R R McCartney 1, M C Schmidt 1
PMCID: PMC1460325  PMID: 9725828

Abstract

The calcium-dependent protein phosphatase calcineurin plays an essential role in ion homeostasis in yeast. In this study, we identify a parallel ion stress response pathway that is independent of the calcineurin signaling pathway. Cells with null alleles in both STD1 and its homologue, MTH1, manifest numerous phenotypes observed in calcineurin mutants, including sodium, lithium, manganese, and hydroxyl ion sensitivity, as well as alpha factor toxicity. Furthermore, increased gene dosage of STD1 suppresses the ion stress phenotypes in calcineurin mutants and confers halotolerance in wild-type cells. However, Std1p functions in a calcineurin-independent ion stress response pathway, since a std1 mth1 mutant is FK506 sensitive under conditions of ion stress. Mutations in other genes known to regulate gene expression in response to changes in glucose concentration, including SNF3, RGT2, and SNF5, also affect cell growth under ion stress conditions. Gene expression studies indicate that the regulation of HAL1 and PMR2 expression is affected by STD1 gene dosage. Taken together, our data demonstrate that response to ion stress requires the participation of both calcineurin-dependent and -independent pathways.

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Selected References

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  1. Albertyn J., Hohmann S., Thevelein J. M., Prior B. A. GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway. Mol Cell Biol. 1994 Jun;14(6):4135–4144. doi: 10.1128/mcb.14.6.4135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alepuz P. M., Cunningham K. W., Estruch F. Glucose repression affects ion homeostasis in yeast through the regulation of the stress-activated ENA1 gene. Mol Microbiol. 1997 Oct;26(1):91–98. doi: 10.1046/j.1365-2958.1997.5531917.x. [DOI] [PubMed] [Google Scholar]
  3. Aperia A., Ibarra F., Svensson L. B., Klee C., Greengard P. Calcineurin mediates alpha-adrenergic stimulation of Na+,K(+)-ATPase activity in renal tubule cells. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7394–7397. doi: 10.1073/pnas.89.16.7394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baldwin S. A. Molecular mechanisms of sugar transport across mammalian and microbial cell membranes. Biotechnol Appl Biochem. 1990 Oct;12(5):512–516. [PubMed] [Google Scholar]
  5. Brewster J. L., de Valoir T., Dwyer N. D., Winter E., Gustin M. C. An osmosensing signal transduction pathway in yeast. Science. 1993 Mar 19;259(5102):1760–1763. doi: 10.1126/science.7681220. [DOI] [PubMed] [Google Scholar]
  6. Carlson M., Osmond B. C., Botstein D. Mutants of yeast defective in sucrose utilization. Genetics. 1981 May;98(1):25–40. doi: 10.1093/genetics/98.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cunningham K. W., Fink G. R. Calcineurin inhibits VCX1-dependent H+/Ca2+ exchange and induces Ca2+ ATPases in Saccharomyces cerevisiae. Mol Cell Biol. 1996 May;16(5):2226–2237. doi: 10.1128/mcb.16.5.2226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cyert M. S., Kunisawa R., Kaim D., Thorner J. Yeast has homologs (CNA1 and CNA2 gene products) of mammalian calcineurin, a calmodulin-regulated phosphoprotein phosphatase. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7376–7380. doi: 10.1073/pnas.88.16.7376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cyert M. S., Thorner J. Regulatory subunit (CNB1 gene product) of yeast Ca2+/calmodulin-dependent phosphoprotein phosphatases is required for adaptation to pheromone. Mol Cell Biol. 1992 Aug;12(8):3460–3469. doi: 10.1128/mcb.12.8.3460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DeRisi J. L., Iyer V. R., Brown P. O. Exploring the metabolic and genetic control of gene expression on a genomic scale. Science. 1997 Oct 24;278(5338):680–686. doi: 10.1126/science.278.5338.680. [DOI] [PubMed] [Google Scholar]
  11. Elledge S. J., Davis R. W. A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae. Gene. 1988 Oct 30;70(2):303–312. doi: 10.1016/0378-1119(88)90202-8. [DOI] [PubMed] [Google Scholar]
  12. Ferrando A., Kron S. J., Rios G., Fink G. R., Serrano R. Regulation of cation transport in Saccharomyces cerevisiae by the salt tolerance gene HAL3. Mol Cell Biol. 1995 Oct;15(10):5470–5481. doi: 10.1128/mcb.15.10.5470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Foor F., Parent S. A., Morin N., Dahl A. M., Ramadan N., Chrebet G., Bostian K. A., Nielsen J. B. Calcineurin mediates inhibition by FK506 and cyclosporin of recovery from alpha-factor arrest in yeast. Nature. 1992 Dec 17;360(6405):682–684. doi: 10.1038/360682a0. [DOI] [PubMed] [Google Scholar]
  14. Ganster R. W., Shen W., Schmidt M. C. Isolation of STD1, a high-copy-number suppressor of a dominant negative mutation in the yeast TATA-binding protein. Mol Cell Biol. 1993 Jun;13(6):3650–3659. doi: 10.1128/mcb.13.6.3650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gietz R. D., Schiestl R. H., Willems A. R., Woods R. A. Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast. 1995 Apr 15;11(4):355–360. doi: 10.1002/yea.320110408. [DOI] [PubMed] [Google Scholar]
  16. Gläser H. U., Thomas D., Gaxiola R., Montrichard F., Surdin-Kerjan Y., Serrano R. Salt tolerance and methionine biosynthesis in Saccharomyces cerevisiae involve a putative phosphatase gene. EMBO J. 1993 Aug;12(8):3105–3110. doi: 10.1002/j.1460-2075.1993.tb05979.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Guerini D. Calcineurin: not just a simple protein phosphatase. Biochem Biophys Res Commun. 1997 Jun 18;235(2):271–275. doi: 10.1006/bbrc.1997.6802. [DOI] [PubMed] [Google Scholar]
  18. Kruckeberg A. L. The hexose transporter family of Saccharomyces cerevisiae. Arch Microbiol. 1996 Nov;166(5):283–292. doi: 10.1007/s002030050385. [DOI] [PubMed] [Google Scholar]
  19. Köhrer K., Domdey H. Preparation of high molecular weight RNA. Methods Enzymol. 1991;194:398–405. doi: 10.1016/0076-6879(91)94030-g. [DOI] [PubMed] [Google Scholar]
  20. Laurent B. C., Treitel M. A., Carlson M. The SNF5 protein of Saccharomyces cerevisiae is a glutamine- and proline-rich transcriptional activator that affects expression of a broad spectrum of genes. Mol Cell Biol. 1990 Nov;10(11):5616–5625. doi: 10.1128/mcb.10.11.5616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liang H., Ko C. H., Herman T., Gaber R. F. Trinucleotide insertions, deletions, and point mutations in glucose transporters confer K+ uptake in Saccharomyces cerevisiae. Mol Cell Biol. 1998 Feb;18(2):926–935. doi: 10.1128/mcb.18.2.926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matheos D. P., Kingsbury T. J., Ahsan U. S., Cunningham K. W. Tcn1p/Crz1p, a calcineurin-dependent transcription factor that differentially regulates gene expression in Saccharomyces cerevisiae. Genes Dev. 1997 Dec 15;11(24):3445–3458. doi: 10.1101/gad.11.24.3445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mendoza I., Rubio F., Rodriguez-Navarro A., Pardo J. M. The protein phosphatase calcineurin is essential for NaCl tolerance of Saccharomyces cerevisiae. J Biol Chem. 1994 Mar 25;269(12):8792–8796. [PubMed] [Google Scholar]
  24. Moser M. J., Geiser J. R., Davis T. N. Ca2+-calmodulin promotes survival of pheromone-induced growth arrest by activation of calcineurin and Ca2+-calmodulin-dependent protein kinase. Mol Cell Biol. 1996 Sep;16(9):4824–4831. doi: 10.1128/mcb.16.9.4824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murguía J. R., Bellés J. M., Serrano R. A salt-sensitive 3'(2'),5'-bisphosphate nucleotidase involved in sulfate activation. Science. 1995 Jan 13;267(5195):232–234. doi: 10.1126/science.7809627. [DOI] [PubMed] [Google Scholar]
  26. Murguía J. R., Bellés J. M., Serrano R. The yeast HAL2 nucleotidase is an in vivo target of salt toxicity. J Biol Chem. 1996 Nov 15;271(46):29029–29033. doi: 10.1074/jbc.271.46.29029. [DOI] [PubMed] [Google Scholar]
  27. Nakamura T., Liu Y., Hirata D., Namba H., Harada S., Hirokawa T., Miyakawa T. Protein phosphatase type 2B (calcineurin)-mediated, FK506-sensitive regulation of intracellular ions in yeast is an important determinant for adaptation to high salt stress conditions. EMBO J. 1993 Nov;12(11):4063–4071. doi: 10.1002/j.1460-2075.1993.tb06090.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Neigeborn L., Carlson M. Genes affecting the regulation of SUC2 gene expression by glucose repression in Saccharomyces cerevisiae. Genetics. 1984 Dec;108(4):845–858. doi: 10.1093/genetics/108.4.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ozcan S., Dover J., Rosenwald A. G., Wölfl S., Johnston M. Two glucose transporters in Saccharomyces cerevisiae are glucose sensors that generate a signal for induction of gene expression. Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12428–12432. doi: 10.1073/pnas.93.22.12428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Posas F., Wurgler-Murphy S. M., Maeda T., Witten E. A., Thai T. C., Saito H. Yeast HOG1 MAP kinase cascade is regulated by a multistep phosphorelay mechanism in the SLN1-YPD1-SSK1 "two-component" osmosensor. Cell. 1996 Sep 20;86(6):865–875. doi: 10.1016/s0092-8674(00)80162-2. [DOI] [PubMed] [Google Scholar]
  31. Pozos T. C., Sekler I., Cyert M. S. The product of HUM1, a novel yeast gene, is required for vacuolar Ca2+/H+ exchange and is related to mammalian Na+/Ca2+ exchangers. Mol Cell Biol. 1996 Jul;16(7):3730–3741. doi: 10.1128/mcb.16.7.3730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stathopoulos A. M., Cyert M. S. Calcineurin acts through the CRZ1/TCN1-encoded transcription factor to regulate gene expression in yeast. Genes Dev. 1997 Dec 15;11(24):3432–3444. doi: 10.1101/gad.11.24.3432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tillman T. S., Ganster R. W., Jiang R., Carlson M., Schmidt M. C. STD1 (MSN3) interacts directly with the TATA-binding protein and modulates transcription of the SUC2 gene of Saccharomyces cerevisiae. Nucleic Acids Res. 1995 Aug 25;23(16):3174–3180. doi: 10.1093/nar/23.16.3174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wach A. PCR-synthesis of marker cassettes with long flanking homology regions for gene disruptions in S. cerevisiae. Yeast. 1996 Mar 15;12(3):259–265. doi: 10.1002/(SICI)1097-0061(19960315)12:3%3C259::AID-YEA901%3E3.0.CO;2-C. [DOI] [PubMed] [Google Scholar]
  37. Wieland J., Nitsche A. M., Strayle J., Steiner H., Rudolph H. K. The PMR2 gene cluster encodes functionally distinct isoforms of a putative Na+ pump in the yeast plasma membrane. EMBO J. 1995 Aug 15;14(16):3870–3882. doi: 10.1002/j.1460-2075.1995.tb00059.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Winston F., Dollard C., Ricupero-Hovasse S. L. Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C. Yeast. 1995 Jan;11(1):53–55. doi: 10.1002/yea.320110107. [DOI] [PubMed] [Google Scholar]
  39. Withee J. L., Mulholland J., Jeng R., Cyert M. S. An essential role of the yeast pheromone-induced Ca2+ signal is to activate calcineurin. Mol Biol Cell. 1997 Feb;8(2):263–277. doi: 10.1091/mbc.8.2.263. [DOI] [PMC free article] [PubMed] [Google Scholar]

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