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Volume 146, Issue 11

A phosphatidylinositol 3-kinase of influences adhesion, filamentous growth and virulence Free

Abstract

To determine if cellular functions of the phosphatidylinositol 3-kinase CaVps34p are related to processes governing pathogenicity, both copies of the gene were sequentially disrupted. Homozygous deletion of resulted in a mutant strain which exhibited defects not only in intracellular vesicle transport processes but also in morphogenesis. The null mutant was unable to form hyphae on different solid media whilst showing a significantly delayed yeast-to-hyphae transition in liquid media. In addition, the mutant was rendered hypersensitive to temperature and osmotic stresses and had a strongly decreased ability to adhere to mouse fibroblast cells compared to the wild-type strain SC5314. Finally, evidence was obtained that is essential for pathogenicity of as the null mutant was shown to be avirulent in a mouse model of systemic infection. pathogenicity was restored to a near wild-type degree upon reintroduction of into the chromosome of the null mutant, demonstrating that the observed avirulence corresponded to the loss of . Thus, the results suggest that may serve as a potential target for antifungal drugs.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Candida albicans , FCS, foetal calf serum , gene disruption , pathogenicity , phosphatidylinositol 3-kinase , PI, phosphatidylinositol and virulence factors
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2000-11-01
2024-04-24
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References

  1. Alex L. A., Korch C., Selitrennikoff C. P., Simon M. I. 1998; COS1, a two component histidine kinase that is involved in hyphal development in the opportunistic pathogen Candida albicans. Proc Natl Acad Sci USA 95:7069–7073 [CrossRef]
     [Google Scholar]
  2. Alonso-Monge R., Navarro-Garcia F., Molero G., Diez-Orejas R., Gustin M., Pla J., Sanchez M., Nombela C. 1999; Role of the mitogen-activated protein kinase Hog1p in morphogenesis and virulence of Candida albicans. J Bacteriol 181:3058–3068
     [Google Scholar]
  3. Boeke J. D., LaCroute F., Fink G. R. 1984; A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet 197:345–346 [CrossRef]
     [Google Scholar]
  4. Borg-von Zepelin M., Wagner T. 1995; Fluorescence assay for the detection of adherent Candida yeasts to target cells in microtest plates. Mycoses 38:339–347 [CrossRef]
     [Google Scholar]
  5. Brown A. J. P., Gow N. A. R. 1999; Regulatory networks controlling Candida albicans morphogenesis. Trends Microbiol 7:333–338 [CrossRef]
     [Google Scholar]
  6. Burd C. G., Emr S. D. 1998; Phosphatidylinositol(3)-phosphate signaling mediated by specific binding to RING FYVE domains. Mol Cell 2:157–162 [CrossRef]
     [Google Scholar]
  7. Cutler J. E. 1991; Putative virulence factors of Candida albicans. Annu Rev Microbiol 45:187–218 [CrossRef]
     [Google Scholar]
  8. DeCamilli P., Emr S. D., McPherson P. S., Novick P. 1996; Phosphoinositides as regulators in membrane traffic. Science 271:1533–1539 [CrossRef]
     [Google Scholar]
  9. Dove S. K., Cooke F. T., Douglas M. R., Sayers L. G., Parker P. J., Mitchell R. H. 1997; Osmotic stress activates phosphatidylinositol-3,5-bisphosphate synthesis. Nature 390:187–192 [CrossRef]
     [Google Scholar]
  10. Eberhard D. A., Cooper C. L., Low M. G., Holz R. W. 1990; Evidence that the inositol phospholipids are necessary for exocytosis. Biochem J 268:15–25
     [Google Scholar]
  11. Eck R., Bergmann C., Ziegelbauer K., Schönfeld W., Künkel W. 1997; A neutral trehalase gene from Candida albicans: molecular cloning, characterization and disruption. Microbiology 143:3747–3756 [CrossRef]
     [Google Scholar]
  12. Eck R., Bruckmann A., Wetzker R., Künkel W. 2000; A phosphatidylinositol 3-kinase of Candida albicans: molecular cloning and characterization. Yeast 16:933–944 [CrossRef]
     [Google Scholar]
  13. Fonzi W. A., Irwin M. Y. 1993; Isogenic strain construction and gene mapping in Candida albicans. Genetics 134:717–728
     [Google Scholar]
  14. Gale C. A., Bendel C. M., McClellan M., Hauser M., Becker J. M., Berman J., Hostetter M. K. 1998; Linkage of adhesion, filamentous growth, and virulence in Candida albicans to a single gene, INT1. Science 279:1355–1358 [CrossRef]
     [Google Scholar]
  15. Gammie A. E., Kurihara L. J., Vallee R. B., Rose M. D. 1995; DNM1, a dynamin-related gene, participates in endosomal trafficking in yeast. J Cell Biol 130:553–566 [CrossRef]
     [Google Scholar]
  16. Gary J. D., Wurmser A. E., Bonangelino C. J., Weisman L. S., Emr S. D. 1998; Fab1p is essential for PtdIns(3)P 5-kinase activity and the maintenance of vacuolar size and membrane homeostasis. J Cell Biol 143:65–79 [CrossRef]
     [Google Scholar]
  17. Herman P. K., Emr S. D. 1990; Characterization of VPS34, a gene required for vacuolar protein sorting and vacuole segregation in Saccharomyces cerevisiae. Mol Cell Biol 10:6742–6754
     [Google Scholar]
  18. Herman P. K., Stack J. H., DeModena J. A., Emr S. D. 1991; A novel protein kinase homolog essential for protein sorting to the yeast lysosome-like vacuole. Cell 64:425–437 [CrossRef]
     [Google Scholar]
  19. Kimura K., Miyake S., Makuuchi M., Morita R., Usui T., Yoshida M., Horinouchi S., Fukui Y. 1995; Phosphatidylinositol 3-kinase in fission yeast: a possible role in stress responses. Biosci Biotechnol Biochem 59:678–682 [CrossRef]
     [Google Scholar]
  20. Köhler J. R., Fink G. R. 1996; Candida albicans strains heterozygous and homozygous for mutations in mitogen-activated protein kinase signaling components have defects in hyphal development. Proc Natl Acad Sci USA 93:13223–13228 [CrossRef]
     [Google Scholar]
  21. Leberer E., Harcus D., Broadbent I. D.7 other authors 1996; Signal transduction through homologs of Ste20p and Ste7p can trigger hyphal formation in the pathogenic fungus Candida albicans. Proc Natl Acad Sci USA 93:13217–13222 [CrossRef]
     [Google Scholar]
  22. Liu H., Köhler J., Fink G. R. 1994; Suppression of hyphal formation in Candida albicans by mutation of a Ste12 homolog. Science 266:1723–1726 [CrossRef]
     [Google Scholar]
  23. Lo H.-J., Köhler J. R., DiDomenico B., Loebenberg D., Cacciapuoti A., Fink G. R. 1997; Nonfilamentous C. albicans mutants are avirulent. Cell 90:939–949 [CrossRef]
     [Google Scholar]
  24. Madhani H. D., Fink G. R. 1998; The control of filamentous differentiation and virulence in fungi. Trends Cell Biol 8:348–353 [CrossRef]
     [Google Scholar]
  25. Munn A. L., Riezman H. 1994; Endocytosis is required for the growth of vacuolar H+-ATPase-defective yeast: identification of six new END genes. J Cell Biol 127:373–386 [CrossRef]
     [Google Scholar]
  26. Navarro-Garcia F., Alonso-Monge R., Rico H., Pla J., Sentandreu R., Nombela C. 1998; A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans. Microbiology 144:411–424 [CrossRef]
     [Google Scholar]
  27. Odds F. C. 1988 Candida and candidosis. A review and bibliography, 2nd edn. London: Bailliere-Tindall;
     [Google Scholar]
  28. Odds F. C. 1994; Pathogenesis of Candida infections. J Am Acad Dermatol 31:S2–S5 [CrossRef]
     [Google Scholar]
  29. Odorizzi G., Babst M., Emr S. D. 1998; Fab1p PtdIns(3)P 5-kinase function essential for protein sorting in the multivesicular body. Cell 95:847–858 [CrossRef]
     [Google Scholar]
  30. Peto R., Pike M. C., Armitage P.7 other authors 1977; Design and analysis of randomized clinical trials requiring prolonged observation of each patient. II. Analysis and examples. Br J Cancer 35:1–39 [CrossRef]
     [Google Scholar]
  31. Piper R. C., Cooper A. A., Yang H., Stevens T. H. 1995; VPS27 controls vacuolar endocytic traffic through a prevacuolar compartment in Saccharomyces cerevisiae. J Cell Biol 131:603–617 [CrossRef]
     [Google Scholar]
  32. Schu P. V., Takegawa K., Fry M. J., Stack J. H., Waterfield M. D., Emr S. D. 1993; Phosphatidylinositol 3-kinase encoded by yeast VPS34 gene essential for protein sorting. Science 260:88–91 [CrossRef]
     [Google Scholar]
  33. Stack J. H., DeWald D. B., Takegawa K., Emr S. D. 1995; Vesicle-mediated protein transport: regulatory interactions between the Vps15 protein kinase and the Vps34 PtdIns 3-kinase essential for protein sorting to the vacuole in yeast. J Cell Biol 129:321–334 [CrossRef]
     [Google Scholar]
  34. Stoldt V. R., Sonneborn A., Leuker C. E., Ernst J. F. 1997; Efg1p, an essential regulator of morphogenesis of the human pathogen Candida albicans, is a member of a conserved class of bHLH proteins regulating morphogenetic processes in fungi. EMBO J 16:1982–1991 [CrossRef]
     [Google Scholar]
  35. Swoboda R. K., Bertram G., Budge S., Gow N. A. R., Gooday G. W., Brown A. J. P. 1995; Structure and regulation of the HSP90 gene from the pathogenic fungus Candida albicans. Infect Immun 63:4506–4514
     [Google Scholar]
  36. Timpel C., Strahl-Bohlsinger S., Ziegelbauer K., Ernst J. F. 1998; Multiple functions of Pmt1p-mediated O-mannosylation in the fungal pathogen Candida albicans. J Biol Chem 273:20837–20846 [CrossRef]
     [Google Scholar]
  37. Toker A., Cantley L. C. 1997; Signalling through the lipid products of phosphoinositide-3-OH kinase. Nature 387:673–676 [CrossRef]
     [Google Scholar]
  38. Vanhaesebroeck B., Leevers S. J., Panayotou G., Waterfield M. D. 1997; Phosphoinositide 3-kinases: a conserved family of signal transducers. Trends Biochem Sci 22:267–272 [CrossRef]
     [Google Scholar]
  39. Weisman L. S., Wicker W. 1992; Molecular characterization of VAC1, a gene required for vacuole inheritance and vacuole protein sorting. J Biol Chem 267:618–623
     [Google Scholar]
  40. Wurmser A. E., Emr S. D. 1998; Phosphoinositide signaling and turnover: PtdIns(3)P, a regulator of membrane traffic, is transported to the vacuole and degraded by a process that requires lumenal vacuolar hydrolase activities. EMBO J 17:4930–4942 [CrossRef]
     [Google Scholar]
  41. Yamamoto A., DeWald D. B., Boronenkov I. V., Anderson R. A., Emr S. D., Koshland D. 1995; Novel PI(4)P 5-kinase homologue, Fab1p, essential for normal vacuole function and morphology in yeast. Mol Biol Cell 6:525–539 [CrossRef]
     [Google Scholar]
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A phosphatidylinositol 3-kinase of Candida albicans influences adhesion, filamentous growth and virulence
Microbiology 146, 2755 (2000); https://doi.org/10.1099/00221287-146-11-2755
/content/journal/micro/10.1099/00221287-146-11-2755
/content/journal/micro/10.1099/00221287-146-11-2755
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