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. 1995 Oct;15(10):5470–5481. doi: 10.1128/mcb.15.10.5470

Regulation of cation transport in Saccharomyces cerevisiae by the salt tolerance gene HAL3.

A Ferrando 1, S J Kron 1, G Rios 1, G R Fink 1, R Serrano 1
PMCID: PMC230797  PMID: 7565698

Abstract

Dynamic regulation of ion transport is essential for homeostasis as cells confront changes in their environment. The gene HAL3 encodes a novel component of this regulatory circuit in the yeast Saccharomyces cerevisiae. Overexpression of HAL3 improves growth of wild-type cells exposed to toxic concentrations of sodium and lithium and suppresses the salt sensitivity conferred by mutation of the calcium-dependent protein phosphatase calcineurin. Null mutants of HAL3 display salt sensitivity. The sequence of HAL3 gives little clue to its function. However, alterations in intracellular cation concentrations associated with changes in HAL3 expression suggest that HAL3 activity may directly increase cytoplasmic K+ and decrease Na+ and Li+. Cation efflux in S. cerevisiae is mediated by the P-type ATPase encoded by the ENA1/PMR24 gene, a putative plasma membrane Na+ pump whose expression is salt induced. Acting in concert with calcineurin, HAL3 is necessary for full activation of ENA1 expression. This functional complementarity is also reflected in the participation of both proteins in recovery from alpha-factor-induced growth arrest. Recently, HAL3 was isolated as a gene (named SIS2) which when overexpressed partially relieves loss of transcription of G1 cyclins in mutants lacking the protein phosphatase Sit4p. Therefore, HAL3 influences cell cycle control and ion homeostasis, acting in parallel to the protein phosphatases Sit4p and calcineurin.

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Selected References

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  1. 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]
  2. 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]
  3. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen P. The structure and regulation of protein phosphatases. Annu Rev Biochem. 1989;58:453–508. doi: 10.1146/annurev.bi.58.070189.002321. [DOI] [PubMed] [Google Scholar]
  8. Cunningham K. W., Fink G. R. Calcineurin-dependent growth control in Saccharomyces cerevisiae mutants lacking PMC1, a homolog of plasma membrane Ca2+ ATPases. J Cell Biol. 1994 Feb;124(3):351–363. doi: 10.1083/jcb.124.3.351. [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. Di Como C. J., Bose R., Arndt K. T. Overexpression of SIS2, which contains an extremely acidic region, increases the expression of SWI4, CLN1 and CLN2 in sit4 mutants. Genetics. 1995 Jan;139(1):95–107. doi: 10.1093/genetics/139.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dmochowska A., Dignard D., Henning D., Thomas D. Y., Bussey H. Yeast KEX1 gene encodes a putative protease with a carboxypeptidase B-like function involved in killer toxin and alpha-factor precursor processing. Cell. 1987 Aug 14;50(4):573–584. doi: 10.1016/0092-8674(87)90030-4. [DOI] [PubMed] [Google Scholar]
  12. Dujon B., Alexandraki D., André B., Ansorge W., Baladron V., Ballesta J. P., Banrevi A., Bolle P. A., Bolotin-Fukuhara M., Bossier P. Complete DNA sequence of yeast chromosome XI. Nature. 1994 Jun 2;369(6479):371–378. doi: 10.1038/369371a0. [DOI] [PubMed] [Google Scholar]
  13. Earnshaw W. C. Anionic regions in nuclear proteins. J Cell Biol. 1987 Oct;105(4):1479–1482. doi: 10.1083/jcb.105.4.1479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Fernandez-Sarabia M. J., Sutton A., Zhong T., Arndt K. T. SIT4 protein phosphatase is required for the normal accumulation of SWI4, CLN1, CLN2, and HCS26 RNAs during late G1. Genes Dev. 1992 Dec;6(12A):2417–2428. doi: 10.1101/gad.6.12a.2417. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gaber R. F., Styles C. A., Fink G. R. TRK1 encodes a plasma membrane protein required for high-affinity potassium transport in Saccharomyces cerevisiae. Mol Cell Biol. 1988 Jul;8(7):2848–2859. doi: 10.1128/mcb.8.7.2848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Galcheva-Gargova Z., Dérijard B., Wu I. H., Davis R. J. An osmosensing signal transduction pathway in mammalian cells. Science. 1994 Aug 5;265(5173):806–808. doi: 10.1126/science.8047888. [DOI] [PubMed] [Google Scholar]
  19. Garciadeblas B., Rubio F., Quintero F. J., Bañuelos M. A., Haro R., Rodríguez-Navarro A. Differential expression of two genes encoding isoforms of the ATPase involved in sodium efflux in Saccharomyces cerevisiae. Mol Gen Genet. 1993 Jan;236(2-3):363–368. doi: 10.1007/BF00277134. [DOI] [PubMed] [Google Scholar]
  20. Gaxiola R., de Larrinoa I. F., Villalba J. M., Serrano R. A novel and conserved salt-induced protein is an important determinant of salt tolerance in yeast. EMBO J. 1992 Sep;11(9):3157–3164. doi: 10.1002/j.1460-2075.1992.tb05392.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Geiser J. R., van Tuinen D., Brockerhoff S. E., Neff M. M., Davis T. N. Can calmodulin function without binding calcium? Cell. 1991 Jun 14;65(6):949–959. doi: 10.1016/0092-8674(91)90547-c. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Guan K. L., Dixon J. E. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem. 1991 Feb 1;192(2):262–267. doi: 10.1016/0003-2697(91)90534-z. [DOI] [PubMed] [Google Scholar]
  24. Gustin M. C., Zhou X. L., Martinac B., Kung C. A mechanosensitive ion channel in the yeast plasma membrane. Science. 1988 Nov 4;242(4879):762–765. doi: 10.1126/science.2460920. [DOI] [PubMed] [Google Scholar]
  25. Haro R., Garciadeblas B., Rodríguez-Navarro A. A novel P-type ATPase from yeast involved in sodium transport. FEBS Lett. 1991 Oct 21;291(2):189–191. doi: 10.1016/0014-5793(91)81280-l. [DOI] [PubMed] [Google Scholar]
  26. Hill J. E., Myers A. M., Koerner T. J., Tzagoloff A. Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast. 1986 Sep;2(3):163–167. doi: 10.1002/yea.320020304. [DOI] [PubMed] [Google Scholar]
  27. Iida H., Yagawa Y., Anraku Y. Essential role for induced Ca2+ influx followed by [Ca2+]i rise in maintaining viability of yeast cells late in the mating pheromone response pathway. A study of [Ca2+]i in single Saccharomyces cerevisiae cells with imaging of fura-2. J Biol Chem. 1990 Aug 5;265(22):13391–13399. [PubMed] [Google Scholar]
  28. 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]
  29. Ko C. H., Gaber R. F. TRK1 and TRK2 encode structurally related K+ transporters in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Aug;11(8):4266–4273. doi: 10.1128/mcb.11.8.4266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kunz J., Hall M. N. Cyclosporin A, FK506 and rapamycin: more than just immunosuppression. Trends Biochem Sci. 1993 Sep;18(9):334–338. doi: 10.1016/0968-0004(93)90069-y. [DOI] [PubMed] [Google Scholar]
  31. Lillie S. H., Brown S. S. Artifactual immunofluorescent labelling in yeast, demonstrated by affinity purification of antibody. Yeast. 1987 Jun;3(2):63–70. doi: 10.1002/yea.320030202. [DOI] [PubMed] [Google Scholar]
  32. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  33. Liu H., Krizek J., Bretscher A. Construction of a GAL1-regulated yeast cDNA expression library and its application to the identification of genes whose overexpression causes lethality in yeast. Genetics. 1992 Nov;132(3):665–673. doi: 10.1093/genetics/132.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Luan S., Li W., Rusnak F., Assmann S. M., Schreiber S. L. Immunosuppressants implicate protein phosphatase regulation of K+ channels in guard cells. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2202–2206. doi: 10.1073/pnas.90.6.2202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lynch J., Polito V. S., Läuchli A. Salinity stress increases cytoplasmic ca activity in maize root protoplasts. Plant Physiol. 1989 Aug;90(4):1271–1274. doi: 10.1104/pp.90.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Marchler G., Schüller C., Adam G., Ruis H. A Saccharomyces cerevisiae UAS element controlled by protein kinase A activates transcription in response to a variety of stress conditions. EMBO J. 1993 May;12(5):1997–2003. doi: 10.1002/j.1460-2075.1993.tb05849.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Morrison D. 14-3-3: modulators of signaling proteins? Science. 1994 Oct 7;266(5182):56–57. doi: 10.1126/science.7939645. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Myers A. M., Tzagoloff A., Kinney D. M., Lusty C. J. Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene. 1986;45(3):299–310. doi: 10.1016/0378-1119(86)90028-4. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  43. Pringle J. R., Adams A. E., Drubin D. G., Haarer B. K. Immunofluorescence methods for yeast. Methods Enzymol. 1991;194:565–602. doi: 10.1016/0076-6879(91)94043-c. [DOI] [PubMed] [Google Scholar]
  44. Ramos J., Alijo R., Haro R., Rodriguez-Navarro A. TRK2 is not a low-affinity potassium transporter in Saccharomyces cerevisiae. J Bacteriol. 1994 Jan;176(1):249–252. doi: 10.1128/jb.176.1.249-252.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Rodriguez-Navarro A., Blatt M. R., Slayman C. L. A potassium-proton symport in Neurospora crassa. J Gen Physiol. 1986 May;87(5):649–674. doi: 10.1085/jgp.87.5.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rose M. D., Broach J. R. Cloning genes by complementation in yeast. Methods Enzymol. 1991;194:195–230. doi: 10.1016/0076-6879(91)94017-7. [DOI] [PubMed] [Google Scholar]
  47. Rudolph H. K., Antebi A., Fink G. R., Buckley C. M., Dorman T. E., LeVitre J., Davidow L. S., Mao J. I., Moir D. T. The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family. Cell. 1989 Jul 14;58(1):133–145. doi: 10.1016/0092-8674(89)90410-8. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Serrano R. H+-ATPase from plasma membranes of Saccharomyces cerevisiae and Avena sativa roots: purification and reconstitution. Methods Enzymol. 1988;157:533–544. doi: 10.1016/0076-6879(88)57102-1. [DOI] [PubMed] [Google Scholar]
  50. Wallis J. W., Chrebet G., Brodsky G., Rolfe M., Rothstein R. A hyper-recombination mutation in S. cerevisiae identifies a novel eukaryotic topoisomerase. Cell. 1989 Jul 28;58(2):409–419. doi: 10.1016/0092-8674(89)90855-6. [DOI] [PubMed] [Google Scholar]
  51. Wiame J. M., Grenson M., Arst H. N., Jr Nitrogen catabolite repression in yeasts and filamentous fungi. Adv Microb Physiol. 1985;26:1–88. doi: 10.1016/s0065-2911(08)60394-x. [DOI] [PubMed] [Google Scholar]
  52. Winston F., Chumley F., Fink G. R. Eviction and transplacement of mutant genes in yeast. Methods Enzymol. 1983;101:211–228. doi: 10.1016/0076-6879(83)01016-2. [DOI] [PubMed] [Google Scholar]
  53. Yazaki P. J., Salvatori S., Sabbadini R. A., Dahms A. S. Calsequestrin, an intracellular calcium-binding protein of skeletal muscle sarcoplasmic reticulum, is homologous to aspartactin, a putative laminin-binding protein of the extracellular matrix. Biochem Biophys Res Commun. 1990 Jan 30;166(2):898–903. doi: 10.1016/0006-291x(90)90895-t. [DOI] [PubMed] [Google Scholar]

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