1887

Abstract

The isolation and functional characterization of a Na/H antiporter gene, is reported here. The gene encodes a protein of 840 amino acids that exhibits high levels of similarity in sequence, size, and structural and functional domains to a group of known Na/H antiporters of fungi. The gene is able to functionally complement the salt-sensitivity of a mutant, and mutations of two conserved aspartate residues to asparagines in the putative Na-binding site abolish this activity. Deletion of results in retardation of growth and a highly elongated morphology in a significant fraction of cells under conditions that normally support yeast growth. These results indicate that has a role in Na and H transport, salt-tolerance, and morphogenesis.

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2000-05-01
2020-09-27
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References

  1. Aronson P. S., Nee J., Suhm M. A.. 1982; Modifier role of internal H+ in activating the Na+–H+ exchanger in renal microvillus membrane vesicles. Nature299:161–163[CrossRef]
    [Google Scholar]
  2. Balcells L., Gomez N., Casamayor A., Clotet J., Arino J.. 1997; Regulation of salt tolerance in fission yeast by a protein-phosphatase-Z-like Ser/Thr protein phosphatase. Eur J Biochem250:476–483[CrossRef]
    [Google Scholar]
  3. Becker D. M., Lundblad V.. 1994; Manipulation of yeast genes. In Current Protocols in Molecular Biology pp.13.7.1–13.7.10Edited by Ausubel F. M..others Chichester: Wiley;
    [Google Scholar]
  4. Beck-Spague C., Jarvis W. R.. 1993; Secular trends in the epidemiology of nosocomial fungal infections in the United States, 1980–1990: national nosocomial infections surveillance system. J Infect Dis167:1247–1251[CrossRef]
    [Google Scholar]
  5. Bernardis F. D., Muhlschlegel F. A., Cassone A., Fonzi W.. 1998; The pH of the host niche controls gene expression in and virulence of Candida albicans. Infect Immun66:3317–3325
    [Google Scholar]
  6. Bianchini L., L’Allemain G., Pouyssegur J.. 1997; The p42/p44 mitogen-activated protein kinase cascade is determinant in mediating activation of the Na+/H+ exchanger (NHE1 isoform) in response to growth factors. J Biol Chem272:271–279[CrossRef]
    [Google Scholar]
  7. Braun B. R., Johnson A. D.. 1997; Control of filament formation in Candida albicans by the transcriptional repressor TUP1. Science277:105–109[CrossRef]
    [Google Scholar]
  8. Cannon R. D., Jenkinson H. F., Shepherd M. G.. 1992; Cloning and expression of Candida albicans ADE2 and proteinase genes on a replicative plasmid in C. albicans and in Saccharomyces cerevisiae. Mol Gen Genet235:453–457[CrossRef]
    [Google Scholar]
  9. Chirgwin T., Przybyla A. E., MacDonald R. J., Rutter W. J.. 1979; Isolation of biologically active ribonucleic acid from sources enriched in ribonucleases. Biochemistry18:5294–5299[CrossRef]
    [Google Scholar]
  10. Cormack B. P., Bertram G., Egerton M., Gow N. A. R., Falkow S., Brown A. J. P.. 1997; Yeast-enhanced green fluorescent protein (yEGFP): a reporter of gene expression in Candida albicans. Microbiology143:303–311[CrossRef]
    [Google Scholar]
  11. Dibrov P., Fliegel L.. 1998; Comparative molecular analysis of Na+/H+ exchangers: a unified model for Na+/H+ antiport. FEBS Lett424:1–5[CrossRef]
    [Google Scholar]
  12. Dibrov P., Smith J. J., Young P. G., Fliegel L.. 1997; Identification and localization of the sod2 gene product in fission yeast. FEBS Lett405:119–124[CrossRef]
    [Google Scholar]
  13. Dibrov P., Young P. G., Fliegel L.. 1998; Functional analysis of amino acid residues essential for activity in the Na+/H+ exchanger of fission yeast. Biochemistry37:8282–8288[CrossRef]
    [Google Scholar]
  14. Fleishman D. G.. 1991; Li+ as a Na+ analog in ion transport process in vertebrates. Tsitologiia33:111–117
    [Google Scholar]
  15. Fonzi W. A., Irwin M. Y.. 1993; Isogenic strain construction and gene mapping in Candida albicans. Genetics134:717–728
    [Google Scholar]
  16. Gow N. A. R.. 1997; Germ tube growth of Candida albicans. Curr Top Med Mycol8:43–55
    [Google Scholar]
  17. Hahnenberger K. M., Jia Z., Young P. G.. 1997; Functional expression of the Scizosaccharomyces pombe Na+/H+ antiporter gene, sod2, in Saccharomyces cerevisiae. Proc Natl Acad Sci USA93:5031–5036
    [Google Scholar]
  18. Hiramatsu T., Kodama K., Kuroda T., Mizushima T., Tsuchiya T.. 1998; A putative multisubunit Na+/H+ antiporter from Staphylococcus aureus. J Bacteriol180:6642–6648
    [Google Scholar]
  19. Inoue H., Noumi T., Tsuchiya T., Kanazawa H.. 1995; Essential aspartic acid residues, Asp-133, Asp-163 and Asp-164, in the transmembrane helices of a Na+/H+ antiporter (NhaA) from Escherichia coli. FEBS Lett363:264–268[CrossRef]
    [Google Scholar]
  20. Iwaki T., Higashida Y., Tsuji H., Tamai Y., Watanabe Y.. 1998; Characterization of a second gene (ZSOD22) of Na+/H+ antiporter from salt-tolerant yeast Zygosaccharomyces rouxii and functional expression of ZSOD2 and ZSOD22 in Saccharomyces cerevisiae. Yeast14:1167–1174[CrossRef]
    [Google Scholar]
  21. Jia Z., McCullough N., Martel R., Hemmingsen S., Young P. G.. 1992; Gene amplification at a locus encoding a putative Na+/H+ antiporter confers sodium and lithium tolerance in fission yeast. EMBO J11:1631–1640
    [Google Scholar]
  22. Kaur S., Mishra P.. 1994; Differential increase in cytoplasmic pH at bud and germ tube formation in Candida albicans: studies of a nongerminative variant. Can J Microbiol40:720–723[CrossRef]
    [Google Scholar]
  23. Kohler G. A., White T. C., Agabian N.. 1997; Overexpression of a cloned IMP dehydrogenase gene of Candida albicans confers resistance to the specific inhibitor mycophenolic acid. J Bacteriol179:2331–2338
    [Google Scholar]
  24. Lee K. L., Buckley H. R., Campbell C. C.. 1975; An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida albicans. Sabouraudia13:148–153[CrossRef]
    [Google Scholar]
  25. Lo H., Kohler J. R., DiDomenico B., Loebenberg D., Cacciapuoti A., Fink G. R.. 1997; Nonfilamentous C. albicans mutants are avirulent. Cell90:939–949[CrossRef]
    [Google Scholar]
  26. Maniatis T., Fritsch E. F., Sambrook J.. 1982; Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  27. Monk B. C., Kurtz M. B., Marrinan J. A., Perlin D. S.. 1991; Cloning and characterization of the plasma membrane H+-ATPase from Candida albicans. J Bacteriol173:6826–6836
    [Google Scholar]
  28. Monk B. C., Niimi M., Shepherd M. G.. 1993; The Candida albicans plasma membrane and H+-ATPase during yeast growth and germ tube formation. J Bacteriol175:5566–5574
    [Google Scholar]
  29. Northrop F. D., Ljubojevic S., Davies J. M.. 1997; Influence of Na+ and anions on the dimorphic transition of Candida albicans. Microbiology143:3757–3765[CrossRef]
    [Google Scholar]
  30. Nozaki K., Kuroda T., Mizushima T., Tsuchiya T.. 1998; A new Na+/H+ antiporter, NhaD, of Vibrio parahaemolyticus. Biochim Biophys Acta1369:213–220[CrossRef]
    [Google Scholar]
  31. Padan E., Schuldiner S.. 1994; Molecular physiology of Na+/H+ antiporters, key transporters in circulation of Na+ and H+ in cells. Biochim Biophys Acta1185:129–151[CrossRef]
    [Google Scholar]
  32. Padan E., Schuldiner S.. 1996; Bacterial Na+/H+ antiporters – molecular biology, biochemistry and physiology. In Handbook of Biological Physicsvol. 2Transport processes in eukaryotic and prokaryotic organisms pp.501–531Edited by Konings W. N., Kaback H. R., Lolkema J. S.. Amsterdam and Oxford: Elsevier;
    [Google Scholar]
  33. Paranjape V., Roy B. G., Datta A.. 1990; Involvement of calcium, calmodulin and phosphorylation in morphogenesis of Candida albicans. J Gen Microbiol136:2149–2154[CrossRef]
    [Google Scholar]
  34. Prior C., Potier S., Souciet J., Sychrova H.. 1996; Characterization of the NHA1 gene encoding a Na+/H+ antiporter of the yeast Saccharomyces cerevisiae. FEBS Lett387:89–93[CrossRef]
    [Google Scholar]
  35. Radford D. R., Challacombe S. J., Walter J. D.. 1994; A scanning electron microscopy investigation of the structure of colonies of different morphologies produced by phenotypic switching of Candida albicans. J Med Microbiol40:416–423[CrossRef]
    [Google Scholar]
  36. Ros R., Montesinos C., Rimon A., Padan E., Serrano R.. 1998; Altered Na+ and Li+ homeostasis in Saccharomyces cerevisiae cells expressing the bacterial cation antiporter NhaA. J Bacteriol180:3131–3136
    [Google Scholar]
  37. Saporito-Irwin S. M., Birse C. E., Syoherd P. S., Fonzi W. A.. 1995; PHR1, a pH-regulated gene of Candida albicans is required for morphogenesis. Mol Cell Biol15:601–613
    [Google Scholar]
  38. Soll D. R.. 1996; The emerging molecular biology of switching in Candida albicans. ASM News62:415–419
    [Google Scholar]
  39. Stewart E., Gow N. A. R., Bowen D. V.. 1988; Cytoplasmic alkalinization during germ tube formation in Candida albicans. J Gen Microbiol134:1079–1087
    [Google Scholar]
  40. Utsugi J., Inaba K., Kuroda T., Tsuda M., Tsuchiya T.. 1998; Cloning and sequencing of a novel Na+/H+ antiporter gene from Pseudomonas aeruginosa. Biochim Biophys Acta1398:330–334[CrossRef]
    [Google Scholar]
  41. Wakabayashi S., Shigekawa M., Pouyssegur J.. 1997; Molecular physiology of vertebrate Na+/H+ exchangers. Physiol Rev77:51–74
    [Google Scholar]
  42. Watanabe Y., Miwa S., Tamai Y.. 1995; Characterization of Na+/H+-antiporter gene closely related to the salt–tolerance of yeast Zygosaccharomyces rouxii. Yeast11:829–838[CrossRef]
    [Google Scholar]
  43. Wieland J., Nitsche A. M., Strayle J., Steiner H., Rudolph H. K.. 1995; The PMR2 gene cluster encodes functionally distinct isoforms of a putative Na+ pump in the yeast plasma membrane. EMBO J14:3870–3882
    [Google Scholar]
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