Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Sep;65(9):3539–3546. doi: 10.1128/iai.65.9.3539-3546.1997

A triple deletion of the secreted aspartyl proteinase genes SAP4, SAP5, and SAP6 of Candida albicans causes attenuated virulence.

D Sanglard 1, B Hube 1, M Monod 1, F C Odds 1, N A Gow 1
PMCID: PMC175504  PMID: 9284117

Abstract

Secreted aspartyl proteinases (Saps) from Candida albicans are encoded by a multigene family with at least nine members (SAP1 to SAP9) and are considered putative virulence factors important for the pathogenicity of this human pathogen. The role of Sap isoenzymes in the virulence of C. albicans has not yet been clearly established, and therefore, using recent progress in the genetics of this yeast, we have constructed a panel of isogenic yeasts, each with a disruption of one or several SAP genes. We focused on the construction of a C. albicans strain in which three related SAP genes (SAP4, SAP5, and SAP6) were disrupted. Growth of the delta sap4,5,6 triple homozygous null mutant DSY459 in complex medium was not affected, whereas, interestingly, growth in a medium containing protein as the sole nitrogen source was severely impaired compared to the growth of the wild-type parent strain SC5314. Since the presence of Sap2 is required for optimal growth on such medium, this suggests that Sap4, Sap5, or Sap6 plays an important role for the process of induction of SAP2. When guinea pigs and mice were injected intravenously with DSY459, their survival time was significantly longer than that of control animals infected with the wild-type SC5314. Attenuated virulence of DSY459 was followed by a significant reduction of yeast cells in infected organs. These data suggest that the group of Sap4, Sap5, and Sap6 isoenzymes is important for the normal progression of systemic infection by C. albicans in animals.

Full Text

The Full Text of this article is available as a PDF (858.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Borg M., Rüchel R. Demonstration of fungal proteinase during phagocytosis of Candida albicans and Candida tropicalis. J Med Vet Mycol. 1990;28(1):3–14. [PubMed] [Google Scholar]
  2. Cutfield S. M., Dodson E. J., Anderson B. F., Moody P. C., Marshall C. J., Sullivan P. A., Cutfield J. F. The crystal structure of a major secreted aspartic proteinase from Candida albicans in complexes with two inhibitors. Structure. 1995 Nov 15;3(11):1261–1271. doi: 10.1016/s0969-2126(01)00261-1. [DOI] [PubMed] [Google Scholar]
  3. Cutler J. E. Putative virulence factors of Candida albicans. Annu Rev Microbiol. 1991;45:187–218. doi: 10.1146/annurev.mi.45.100191.001155. [DOI] [PubMed] [Google Scholar]
  4. De Bernardis F., Cassone A., Sturtevant J., Calderone R. Expression of Candida albicans SAP1 and SAP2 in experimental vaginitis. Infect Immun. 1995 May;63(5):1887–1892. doi: 10.1128/iai.63.5.1887-1892.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. De Bernardis F., Chiani P., Ciccozzi M., Pellegrini G., Ceddia T., D'Offizzi G., Quinti I., Sullivan P. A., Cassone A. Elevated aspartic proteinase secretion and experimental pathogenicity of Candida albicans isolates from oral cavities of subjects infected with human immunodeficiency virus. Infect Immun. 1996 Feb;64(2):466–471. doi: 10.1128/iai.64.2.466-471.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  7. Fonzi W. A., Irwin M. Y. Isogenic strain construction and gene mapping in Candida albicans. Genetics. 1993 Jul;134(3):717–728. doi: 10.1093/genetics/134.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fusek M., Smith E. A., Monod M., Dunn B. M., Foundling S. I. Extracellular aspartic proteinases from Candida albicans, Candida tropicalis, and Candida parapsilosis yeasts differ substantially in their specificities. Biochemistry. 1994 Aug 16;33(32):9791–9799. doi: 10.1021/bi00198a051. [DOI] [PubMed] [Google Scholar]
  9. Hube B., Monod M., Schofield D. A., Brown A. J., Gow N. A. Expression of seven members of the gene family encoding secretory aspartyl proteinases in Candida albicans. Mol Microbiol. 1994 Oct;14(1):87–99. doi: 10.1111/j.1365-2958.1994.tb01269.x. [DOI] [PubMed] [Google Scholar]
  10. Hube B., Sanglard D., Odds F. C., Hess D., Monod M., Schäfer W., Brown A. J., Gow N. A. Disruption of each of the secreted aspartyl proteinase genes SAP1, SAP2, and SAP3 of Candida albicans attenuates virulence. Infect Immun. 1997 Sep;65(9):3529–3538. doi: 10.1128/iai.65.9.3529-3538.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ibrahim A. S., Mirbod F., Filler S. G., Banno Y., Cole G. T., Kitajima Y., Edwards J. E., Jr, Nozawa Y., Ghannoum M. A. Evidence implicating phospholipase as a virulence factor of Candida albicans. Infect Immun. 1995 May;63(5):1993–1998. doi: 10.1128/iai.63.5.1993-1998.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jaton-Ogay K., Paris S., Huerre M., Quadroni M., Falchetto R., Togni G., Latgé J. P., Monod M. Cloning and disruption of the gene encoding an extracellular metalloprotease of Aspergillus fumigatus. Mol Microbiol. 1994 Dec;14(5):917–928. doi: 10.1111/j.1365-2958.1994.tb01327.x. [DOI] [PubMed] [Google Scholar]
  13. Kirsch D. R., Whitney R. R. Pathogenicity of Candida albicans auxotrophic mutants in experimental infections. Infect Immun. 1991 Sep;59(9):3297–3300. doi: 10.1128/iai.59.9.3297-3300.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kwon-Chung K. J., Lehman D., Good C., Magee P. T. Genetic evidence for role of extracellular proteinase in virulence of Candida albicans. Infect Immun. 1985 Sep;49(3):571–575. doi: 10.1128/iai.49.3.571-575.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Macdonald F., Odds F. C. Inducible proteinase of Candida albicans in diagnostic serology and in the pathogenesis of systemic candidosis. J Med Microbiol. 1980 Aug;13(3):423–435. doi: 10.1099/00222615-13-3-423. [DOI] [PubMed] [Google Scholar]
  16. Macdonald F., Odds F. C. Virulence for mice of a proteinase-secreting strain of Candida albicans and a proteinase-deficient mutant. J Gen Microbiol. 1983 Feb;129(2):431–438. doi: 10.1099/00221287-129-2-431. [DOI] [PubMed] [Google Scholar]
  17. Monod M., Paris S., Sarfati J., Jaton-Ogay K., Ave P., Latgé J. P. Virulence of alkaline protease-deficient mutants of Aspergillus fumigatus. FEMS Microbiol Lett. 1993 Jan 1;106(1):39–46. doi: 10.1111/j.1574-6968.1993.tb05932.x. [DOI] [PubMed] [Google Scholar]
  18. Monod M., Togni G., Hube B., Sanglard D. Multiplicity of genes encoding secreted aspartic proteinases in Candida species. Mol Microbiol. 1994 Jul;13(2):357–368. doi: 10.1111/j.1365-2958.1994.tb00429.x. [DOI] [PubMed] [Google Scholar]
  19. Odds F. C. Antifungal susceptibility testing of Candida spp. by relative growth measurement at single concentrations of antifungal agents. Antimicrob Agents Chemother. 1992 Aug;36(8):1727–1737. doi: 10.1128/aac.36.8.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ollert M. W., Söhnchen R., Korting H. C., Ollert U., Bräutigam S., Bräutigam W. Mechanisms of adherence of Candida albicans to cultured human epidermal keratinocytes. Infect Immun. 1993 Nov;61(11):4560–4568. doi: 10.1128/iai.61.11.4560-4568.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ray T. L., Payne C. D. Scanning electron microscopy of epidermal adherence and cavitation in murine candidiasis: a role for Candida acid proteinase. Infect Immun. 1988 Aug;56(8):1942–1949. doi: 10.1128/iai.56.8.1942-1949.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reichard U., Monod M., Odds F., Rüchel R. Virulence of an aspergillopepsin-deficient mutant of Aspergillus fumigatus and evidence for another aspartic proteinase linked to the fungal cell wall. J Med Vet Mycol. 1997 May-Jun;35(3):189–196. doi: 10.1080/02681219780001131. [DOI] [PubMed] [Google Scholar]
  23. Ross I. K., De Bernardis F., Emerson G. W., Cassone A., Sullivan P. A. The secreted aspartate proteinase of Candida albicans: physiology of secretion and virulence of a proteinase-deficient mutant. J Gen Microbiol. 1990 Apr;136(4):687–694. doi: 10.1099/00221287-136-4-687. [DOI] [PubMed] [Google Scholar]
  24. Rüchel R., Böning-Stutzer B., Mari A. A synoptical approach to the diagnosis of candidosis, relying on serological antigen and antibody tests, on culture, and on evaluation of clinical data. Mycoses. 1988 Feb;31(2):87–106. [PubMed] [Google Scholar]
  25. Rüchel R., Ritter B., Schaffrinski M. Modulation of experimental systemic murine candidosis by intravenous pepstatin. Zentralbl Bakteriol. 1990 Aug;273(3):391–403. doi: 10.1016/s0934-8840(11)80443-3. [DOI] [PubMed] [Google Scholar]
  26. Rüchel R., Zimmermann F., Böning-Stutzer B., Helmchen U. Candidiasis visualised by proteinase-directed immunofluorescence. Virchows Arch A Pathol Anat Histopathol. 1991;419(3):199–202. doi: 10.1007/BF01626348. [DOI] [PubMed] [Google Scholar]
  27. Sanglard D., Fiechter A. DNA transformations of Candida tropicalis with replicating and integrative vectors. Yeast. 1992 Dec;8(12):1065–1075. doi: 10.1002/yea.320081209. [DOI] [PubMed] [Google Scholar]
  28. Sanglard D., Ischer F., Monod M., Bille J. Susceptibilities of Candida albicans multidrug transporter mutants to various antifungal agents and other metabolic inhibitors. Antimicrob Agents Chemother. 1996 Oct;40(10):2300–2305. doi: 10.1128/aac.40.10.2300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanglard D., Togni G., de Viragh P. A., Monod M. Disruption of the gene encoding the secreted acid protease (ACP) in the yeast Candida tropicalis. FEMS Microbiol Lett. 1992 Aug 15;74(2-3):149–156. doi: 10.1016/0378-1097(92)90421-j. [DOI] [PubMed] [Google Scholar]
  30. Shepherd M. G. Pathogenicity of morphological and auxotrophic mutants of Candida albicans in experimental infections. Infect Immun. 1985 Nov;50(2):541–544. doi: 10.1128/iai.50.2.541-544.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Togni G., Sanglard D., Falchetto R., Monod M. Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis. FEBS Lett. 1991 Jul 29;286(1-2):181–185. doi: 10.1016/0014-5793(91)80969-a. [DOI] [PubMed] [Google Scholar]
  32. Togni G., Sanglard D., Monod M. Acid proteinase secreted by Candida tropicalis: virulence in mice of a proteinase negative mutant. J Med Vet Mycol. 1994;32(4):257–265. doi: 10.1080/02681219480000331. [DOI] [PubMed] [Google Scholar]
  33. White T. C., Agabian N. Candida albicans secreted aspartyl proteinases: isoenzyme pattern is determined by cell type, and levels are determined by environmental factors. J Bacteriol. 1995 Sep;177(18):5215–5221. doi: 10.1128/jb.177.18.5215-5221.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES