1887

Abstract

A system for controlling gene expression was established in the pathogenic fungus Candida glabrata to elucidate the physiological functions of genes. To control the expression of the gene of interest, the cells were first transformed with the plasmid carrying the tetracycline repressor-transactivator fusion then with the DNA fragment containing the controllable cassette, the tetracycline operator chimeric promoter The peptide elongation factor 3 and DNA topoisomerase II genes from were cloned and their expression assessed using this system. When the promoter of or was replaced with doxycycline almost completely repressed the expression of both mRNAs, and impaired growth. Repression of the or gene by doxycycline also hampered the survival of cells in mice; in mouse kidneys the number of cells, in which the or promoter was replaced with the controllable cassette, did not increase when the mice were given doxycycline. Thus, it appears that the gene repression mediated by doxycycline occurred not only in culture media but also in animals; therefore, this system can be used to elucidate the function of the gene in fungal infections and pathogenesis.

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1998-09-01
2021-10-16
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References

  1. Aisner J., Schimpff S.C., Sutherland J.C., Young V.M., Wiernik P.H. 1976; Torulopsis glabrata infections in patients with cancer: increasing incidence and relationship to colonization.. Am J Med 61:23–28
    [Google Scholar]
  2. Barns S.M., Lane D.J., Sogin M.L., Bibeau C., Weisburg W.G. 1991; Evolutionary relationships among pathogenic Candida species and relatives.. J Bacterial 173:2250–2255
    [Google Scholar]
  3. DiNardo S., Voelkel K., Sternglanz R. 1984; DNA topo-isomerase II mutant of Saccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication.. Proc Natl Acad Sci USA 81:2616–2620
    [Google Scholar]
  4. Di Domenico B.J., Lupisella J., Sandbaken M., Chakraburtty K. 1992; Isolation and sequence analysis of the gene encoding translation elongation factor 3 from Candida albicans. . Yeast 8:337–352
    [Google Scholar]
  5. Figgitt D.P., Denyer S.P., Dewick P.M., Jackson D.E., Williams P. 1989; Topoisomerase II: a potential target for novel antifungal agent.. Biochem Biophys Res Commun 160:257–262
    [Google Scholar]
  6. Gallis H.A., Drew R.H., Pickard W.W. 1990; Amphotericin B: 30 years of clinical experience.. Rev Infect Dis 12:308–329
    [Google Scholar]
  7. Georgopapadakou N.H., Walsh T.J. 1996; Antifungal agents: chemotherapeutic targets and immunologic strategies.. Anti- microb Agents Chemother 40:279–291
    [Google Scholar]
  8. Giaever G., Lynn R., Goto T., Wang J.C. 1986; The complete nucleotide sequence of the structural gene TOP2 of yeast DNA topoisomerase II.. J Biol Chem 261:12448–12454
    [Google Scholar]
  9. Goto T., Wang J.C. 1984; Yeast DNA topoisomerase II is encoded by a single-copy, essential gene.. Cell 36:1073–1080
    [Google Scholar]
  10. Hickey W.F., Sommerville L.H., Schoen F.J. 1983; Dis-seminated Candida glabrata - report of a uniquely severe inäfection and a literature review.. Am J Clin Pathol 80:724–727
    [Google Scholar]
  11. Hitchcock C.A., Pye G.W., Troke P.F., Johnson E.M., Warnock D.W. 1993; Fluconazole resistance in Candida glabrata. . Antimicrob Agents Chemother 37:1962–1965
    [Google Scholar]
  12. Holm C., Goto T., Wang J.C., Botstein D. 1985; DNA topoisomerase II is required at the time of mitosis in yeast.. Cell 41:553–563
    [Google Scholar]
  13. Jenkins J.R., Ayton P., Jones T., Davies S.L., Simmons D.L., Harris A.L., Sheer D., Hickson I.D. 1992; Isolation of cDNA clones encoding the beta isozyme of human DNA topoisomerase II and localization of the gene to chromosome 3p24.. Nucleic Acids Res 20:5587–5592
    [Google Scholar]
  14. Kitada K., Yamaguchi E., Arisawa M. 1995; Cloning of the Candida glabrata TRP1 and HIS3 genes, and construction of their disruptant strains by sequential integrative transformation.. Gene 165:203–206
    [Google Scholar]
  15. Keller B.A., Patel S., Fisher L.M. 1997; Molecular cloning and expression of the Candida albicans TOP2 gene allows study of fungal DNA topoisomerase II inhibitors in yeast.. Biochem J 324:329–339
    [Google Scholar]
  16. Myers K.K., Fonzi W.A., Sypherd P.S. 1992; Isolation and sequence analysis of the gene for translation elongation factor 3 from Candida albicans. . Nucleic Acids Res 20:1705–1710
    [Google Scholar]
  17. Nagahashi S., Nakayama H., Hamada K., Yang H., Arisawa M., Kitada K. 1997; Regulation by tetracycline of gene expression in Saccharomyces cerevisiae. . Mol Gen Genet 273:372–375
    [Google Scholar]
  18. Newman S.L., Flanigan T.P., Fisher A., Rinaldi M.G., Stein M., Vigilante K. 1994; Clinically significant mucosal candidiasis resistant to fluconazole treatment in patients with AIDS.. Clin Infect Dis 19:684–686
    [Google Scholar]
  19. Qin S.L., Xie A.G., Bonato M.C., McLaughlin C.S. 1990; Sequence analysis of the translation elongation factor 3 from Saccharomyces cerevisiae. . J Biol Chem 265:1903–1912
    [Google Scholar]
  20. Rex J.H., Rinaldi M.G., Pfaller M.A. 1995; Resistance of Candida species to fluconazole.. Antimicrob Agents Chemother 39:1–8
    [Google Scholar]
  21. Rose M.D., Winston F., Hieter P. 1990 Methods in Yeast Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  22. Sandbaken M.G., Lupisella J.A., DiDomenico B., Chakraburtty K. 1990; Protein synthesis in yeast. Structural and functional analysis of the gene encoding elongation factor 3.. J Biol Chem 265:15838–15844
    [Google Scholar]
  23. Shen L.L., Baranowski J., Fostel J., Montgomery D.A., Lartey P.A. 1992; DNA topoisomerases from pathogenic fungi: targets for the discovery of antifungal drugs.. Antimicrob Agents Chemother 36:2778–2784
    [Google Scholar]
  24. Triana-Alonso F.J., Chakraburtty K., Nierhaus K.H. 1995; The elongation factor 3 unique in higher fungi and essential for protein biosynthesis is an E site factor.. J Biol Chem 270:20473–20478
    [Google Scholar]
  25. Tsai-Pflugfelder M., Liu L.F., Liu A.A., Tewey K.M., Whang-Peng J., Knutsen T., Huebner K., Croce C.M., Wang J.C. 1988; Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22.. Proc Natl Acad Sci USA 85:7177–7181
    [Google Scholar]
  26. Uemura T., Yanagida M. 1986; Mitotic spindle pulls but fails to separate chromosomes in type II DNA topoisomerase mutants: uncoordinated mitosis.. EMBO J 5:1003–1010
    [Google Scholar]
  27. Uemura T., Morikawa K., Yanagida M. 1986; The nucleotide sequence of the fission yeast DNA topoisomerase II gene: structural and functional relationships to other DNA topoisomerases.. EMBO J 5:2355–2361
    [Google Scholar]
  28. Vershon A.K., Hollingsworth N.M., Johnston A.D. 1992; Meiotic induction of the yeast HOPI gene is controlled by positive and negative regulatory sites.. Mol Cell Biol 12:3706–3714
    [Google Scholar]
  29. Wingard J.R. 1995; Importance of Candida species other than C. albicans as pathogens in oncology patients.. Clin Infec Dis 115:115–125
    [Google Scholar]
  30. Worland S.T., Wang J.C. 1989; Inducible overexpression, purification, and active site mapping of DNA topoisomerase II from the yeast Saccharomyces cerevisiae. . J Biol Chem 264:4412–1416
    [Google Scholar]
  31. Wyckoff E., Natalie D., Nolan J.M., Lee M., Hsieh T. 1989; Structure of the Drosophila DNA topoisomerase II gene. Nucleoätide sequence and homology among topoisomerases II.. J Mol Biol 205:1–13
    [Google Scholar]
  32. Ypma-Wong M.F., Fonzi W.A., Sypherd P.S. 1992; Fungus-specific translation elongation factor 3 gene present in Pneumo- cytis carinii. . Infect Immun 60:4140–4145
    [Google Scholar]
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