1887

Abstract

The virulence of strains deficient in fatty acid synthase activity by virtue of disruption/deletion of the gene was examined in a rat model of oropharyngeal candidiasis. The alleles of CA14 (∆) were sequentially disrupted with a cassette that included a portion of from which a 984 bp fragment containing the FAS condensing reaction domain was deleted and replaced with sequences. Verification of fatty acid synthase inactivation was obtained from assays of enzyme activity. Strains in which a single allele was disrupted (CFD1 and CFD3) exhibited an approximately 20% reduction in activity, when compared to wild-type. In addition, fatty acid synthase activity was abolished in a null mutant strain (CFD2), and growth of CFD2 occurred only when the growth medium was supplemented with Tween 40 and certain fatty acids. Strain CFD2 was avirulent in the rat model, indicating that fatty acid synthase activity is required for oropharyngeal infection. Strains with a single allele disruption colonized the oral cavity, but the number of cells recovered from infected animals was approximately fivefold less than for the parental strain. The results suggest that FAS may be exploited as a possible target for the development of new antifungal agents.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-142-9-2509
1996-09-01
2021-05-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/9/mic-142-9-2509.html?itemId=/content/journal/micro/10.1099/00221287-142-9-2509&mimeType=html&fmt=ahah

References

  1. Becker J. M., Henry L. K., Jiang W., Koltin Y. 1995; Reduced virulence of Candida albicans mutants affected in multidrug resistance. Infect Immun 63:4528–4530
    [Google Scholar]
  2. Boeke J. D., Lacroute F., Fink G. R. 1984; A positive selection tor mutants lacking orotidine-5-phosphate decarboxylase activity in yeast: 5 fluoro-orotic acid resistance. Mol Gen Genet 197:345–346
    [Google Scholar]
  3. Broedel S. E. Jr Zhao X.-J., Cihlar R. L. 1996; Fatty acid FL44 synthase as a target in the development of new antifungals. Recent Res Dev Antimicrob Agents Chemother 1:25–33
    [Google Scholar]
  4. Bulawa C. E., Miller D. W., Henry LK., Becker J. M. 1995; Attenuated virulence of chitin-deficient mutants ofCandida albicans . Proc Natl Acad Sci USA 92:10570–10574
    [Google Scholar]
  5. Cole M. F., Bowen W. H., Zhao X.-J., Cihlar R. L. 1995; Avirulence ol Candida albicans auxotrophic mutants in a rat model of oropharyngeal candidiasis. FEMS Microbiol Lett 126:177–180
    [Google Scholar]
  6. Cutler J. E. 1991; Putative virulence factors of Candida albicans . Annu Rev Microbiol 45:187–218
    [Google Scholar]
  7. De Bernardis F., Adriani D., Lorenzini R., Pontieu E., Carruba G., Cassone A. 1993; Filamentous growth and elevated vagmo pathic potential of a nongerminative variant of Candida albicans expressing low virulence in systemic infection. Infect Immun 61:1500–1508
    [Google Scholar]
  8. Fonzi W., Irwin M. Y. 1993; Isogenic strain construction and gene mapping inCandida albicans . Genetics 134:717–728
    [Google Scholar]
  9. Ghannoum M. A., Spellberg B., Saporito-lrwin S. M., Fonzi W. F. 1995; Reduced virulence of Candida albicans PHR1 mutants. Infect Immun 63:4528–4530
    [Google Scholar]
  10. Gillum A. M., Tsay W. Y. H., Kirsch D. R. 1984; Isolation of the Candida albicans gene for orotidine-5´-phosphate decarboxylase by complementation of 3.cerevisiae ura3 and E. coli pyrF mutations. Mol Gen Genet 198:179–182
    [Google Scholar]
  11. Holmberg K., Meyer R. D. 1986; Fungal infections in patients with AIDS and AIDS-related complex. Scand j Infect Dis 18:179–185
    [Google Scholar]
  12. Kuziora M. A., Chalmer J. H. Jr Douglas M. G., Hitzeman R. A., Mattick J. S., Wakil S. J. 1983; Molecular cloning ol fatty acid synthetase genes from Saccharomyces cerevisiae . J Biol Gbern 258:11648–11653
    [Google Scholar]
  13. Lynen F. 1969; Yeast fatty acid synthase. Methods Enzymol 14:17–33
    [Google Scholar]
  14. McElhaney-Feser G. E., Cihlar R. L. 1994; Purification and characterization of fatty acid synthase from Candida albicans, strain 4918, and two derived sponraneous cerulenin-resistant mutants. J Med Vet Mycol 32:13–20
    [Google Scholar]
  15. Meitner S. W., Bowen W. H., Haidaris C.G. 1990; Oral and esophageal Candida albicans infection in hyposalivatory rats. Infect Immun 58:2228–2236
    [Google Scholar]
  16. Odds F. C. 1988 Candida and Candidosis. London: Baillière Tindall;
    [Google Scholar]
  17. Ōmura S. 1981; Cerulenin. Methods Enzymol 72:520–532
    [Google Scholar]
  18. Ruchel R., De Bernardis F., Ray T. L., Sullivan P. A., Cole G. T. 1992; Candida acid proteinases. J Med Vet Mycol 29: (Suppl.) 1–9
    [Google Scholar]
  19. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molectdar Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Schweizer E., Müller G., Roberts L. M., Schweizer M., Rösch L., Weisner P., Beck J., Stratman D., Zauner I. 1987; Genetic control of fatty acid synthetase biosynthesis and structure in lower fungi. Fat Sci Technol 89:570–577
    [Google Scholar]
  21. Sherman F., Fink G. R., Hicks J. B. 1986 Laboratory Course Manualfor Methods in Yeast Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  22. Siggaard-Anderson M. 1993; Conserved residues in condensing enzyme domains of fatty acid synthases and related sequences. Protein Seq Data Anal 5:325–335
    [Google Scholar]
  23. Smith S. 1994; The animal fatty acid synthase: one gene, one polypeptide, seven enzymes. FASEB J 8:1248–1259
    [Google Scholar]
  24. Soll D. R. 1992; High frequency switching in Candida albicans . Clin Microbiol Rev 5:183–203
    [Google Scholar]
  25. Stoops J. K., Awad E. S., Arslanian M. J., Gussberg S., Wakil S. J. 1978; Studies on the yeast fatty acid synthetase: subunit composition and structural organization of a large multifunctional enzyme complex. J Biol Chem 253:4464–4475
    [Google Scholar]
  26. Southard S. B., Cihlar R. L. 1995; Analysis and expression of the Candida albicans FAS2 gene. Gene 156:133–138
    [Google Scholar]
  27. Witkowski A., Rangan V. S., Randhawa Z. I., Amy C. M., Smith S. 1991; Structural organization of the multifunctional animal fatty-acid synthase. Eur J Biochem 198:571–579
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-142-9-2509
Loading
/content/journal/micro/10.1099/00221287-142-9-2509
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error