1887

Abstract

We report here the cloning of the genomic topoisomerase I gene () by use of PCR and subsequent hybridization. The predicted protein sequence shared 58.8% identity with the topoisomerase I and 30-50% identity with other eukaryotic topoisomerase I proteins. A conditional gene disruption strain (CWJ477) was constructed so that one copy of was deleted and the other copy of was placed under a regulatable promoter. Under repressed conditions, cells grew slowly and cell morphology was abnormal. The virulence of CWJ477 was markedly reduced in a mouse model system, and that of the single gene knockout strain was slightly attenuated, indicating that might play a role in the infection of in mice in a dose-dependent manner. Despite the reduced virulence of both the single and double knockout strains, viable cells of the pathogen were recovered from the kidneys as late as 22 d post-infection.

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/content/journal/micro/10.1099/00221287-143-2-377
1997-02-01
2024-04-18
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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 63:4515–4518
    [Google Scholar]
  2. Bulawa C.E., Miller D.W., Henry L.K., Becker J.M. 1995; Attenuated virulence of chitin-deficient mutants of Candida albicans. . Proc Natl Acad Sci USA 9210570–10574
    [Google Scholar]
  3. Chen A.Y., Liu L.F. 1994; DNA topoisomerases: essential enzymes and lethal targets.. Annu Rev Pharmacol Toxicol 34:191–218
    [Google Scholar]
  4. Christman M.F., Dietrich F.S., Fink G.R. 1988; Mitotic recombination in the rDNA of S. cerevisiae is suppressed by the combined action of DNA topoisomerases I and II.. Cell 55:413–425
    [Google Scholar]
  5. Fonzi W.A., Irwin M.Y. 1993; Isogenic strain construction and gene mapplng in Candida albicans. . Genetics 134:717–728
    [Google Scholar]
  6. Fostel J., Montgomery D. 1995; Identification of the aminocatechol A-3253 as an in vitro poison of DNA topo- isomerase I from Candida albicans. . Antimicrob Agents Chemother 39:586–592
    [Google Scholar]
  7. Fostel J.M., Montgomery D.A., Shen L.L. 1992; Characterization of DNA topoisomerase I from Candida albicans as a target for drug discovery.. Antimicrob Agents Chemother 36:2131–2138
    [Google Scholar]
  8. Gerhold D"Thiyagarajan, Kmiec E.B. 1994; The topoisomerase I gene from Ustilago maydis: sequence, disruption and mutant phenotype.. Nucleic Acids Res 22:3773–3778
    [Google Scholar]
  9. Goto T., Wang J.C. 1985; Cloning of yeast TOP1, the gene encoding DNA topoisomerase I, and construction of mutants defective in both DNA topoisomerase I and DNA topoisomerase II.. Proc Natl Acad Sci USA 827178–7182
    [Google Scholar]
  10. Kirsch D.R., Whitney R.R. 1991; Pathogenicity of Candida albicans auxotrophic mutants in experimental infections.. Infect Immun 59:3297–3300
    [Google Scholar]
  11. Kwon-Chung K.J., Bennett J.E. 1992; Candidiasis.. In Medical Mycology pp. 280–336 Philadelphia:: Lea Febiger.;
    [Google Scholar]
  12. Lee M.P., Brown S.D., Chen A., Hsieh T.S. 1993; DNAtopoisomerase I is essential in Drosophila melanogaster. . Proc Natl Acad Sci USA 906656–6660
    [Google Scholar]
  13. Levin N.A., Bjornsti M.A., Fink G.R. 1993; A novel mutation in DNA topoisomerase I of yeast causes DNA damage and RAD9-dependent cell cycle arrest.. Genetics 133:799–814
    [Google Scholar]
  14. Omaha T., Suzuki T., Mori M., Osawa S., Ueda T., Watanabe K., Nakase T. 1993; Non-universal decoding of the leucine codon CUG in several Candida species.. Nucleic Acids Res 21:4039–4045
    [Google Scholar]
  15. Roca J. 1995; The mechanisms of DNA topoisomerases.. Trends Biochem Sci 20:156–160
    [Google Scholar]
  16. Rosenbluh A., Mevarech M., Koltin Y., Gorman J.A. 1985; Isolation of genes from Candida albicans by complementation in Saccharomyces cerevisiae. . Mol Gen Genet 200:500–502
    [Google Scholar]
  17. Thrash C., Bankier A.T., Barrell B.G., Sternglanz R. 1985; Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene.. Proc Natl Acad Sci USA 824374–4378
    [Google Scholar]
  18. Uemura T., Morino K., Uzawa S., Shiozaki K., Yanagida M. 1987; Cloning and sequencing of Schizosaccharomyces pombe DNA topoisomerase I gene, and effect of gene disruption.. Nucleic Acids Res 15:9727–9739
    [Google Scholar]
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