1887

Abstract

One of the mediators of pleiotropic drug resistance in is the ABC-transporter gene . This gene is regulated by at least two transcription factors with Zn(2)-Cys(6) finger DNA-binding motifs, Pdr1p and Pdr3p. In this work, we searched for functional homologues of these transcription factors in . A gene library was screened in a mutant lacking and and clones resistant to azole antifungals were isolated. From these clones, three genes responsible for azole resistance were identified. These genes (, and ) encode proteins with Zn(2)-Cys(6)-type zinc finger motifs in their N-terminal domains. The genes expressed in could activate the transcription of a - reporter system and this reporter activity was PDRE-dependent. They could also confer resistance to azoles in a strain lacking , and , suggesting that , and dependent azole resistance can be caused by genes other than in . Deletion of , and in had no effect on fluconazole susceptibility and did not alter the expression of the ABC-transporter genes and or the major facilitator gene , which encode multidrug transporters known as mediators of azole resistance in . However, additional phenotypic screening tests on the mutants revealed that the presence of was necessary to sustain growth on non-fermentative carbon sources (sodium acetate, acetic acid, ethanol). In conclusion, possesses functional homologues of the Pdr1p and Pdr3p transcription factors; however, their properties in have been rewired to other functions.

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2008-05-01
2019-10-17
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References

  1. Akache, B., Wu, K. & Turcotte, B. ( 2001; ). Phenotypic analysis of genes encoding yeast zinc cluster proteins. Nucleic Acids Res 29, 2181–2190.[CrossRef]
    [Google Scholar]
  2. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K. ( 1987; ). Current Protocols in Molecular Biology: Greene Publishing Associates and Wiley-Interscience.
  3. Balzi, E., Wang, M., Leterme, S., Van Dyck, L. & Goffeau, A. ( 1994; ). PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1. J Biol Chem 269, 2206–2214.
    [Google Scholar]
  4. Brand, A., MacCallum, D. M., Brown, A. J., Gow, N. A. & Odds, F. C. ( 2004; ). Ectopic expression of URA3 can influence the virulence phenotypes and proteome of Candida albicans but can be overcome by targeted reintegration of URA3 at the RPS10 locus. Eukaryot Cell 3, 900–909.[CrossRef]
    [Google Scholar]
  5. Carvajal, E., van den Hazel, H. B., Cybularz-Kolaczkowska, A., Balzi, E. & Goffeau, A. ( 1997; ). Molecular and phenotypic characterization of yeast PDR1 mutants that show hyperactive transcription of various ABC multidrug transporter genes. Mol Gen Genet 256, 406–415.[CrossRef]
    [Google Scholar]
  6. Chiranand, W., McLeod, I., Zhou, H., Lynn, J. J., Vega, L. A., Myers, H., Yates, J. R., III, Lorenz, M. C. & Gustin, M. C. ( 2008; ). CTA4 transcription factor mediates induction of nitrosative stress response in Candida albicans. Eukaryot Cell 7, 268–278.[CrossRef]
    [Google Scholar]
  7. Coste, A. T., Karababa, M., Ischer, F., Bille, J. & Sanglard, D. ( 2004; ). TAC1, transcriptional activator of CDR genes, is a new transcription factor involved in the regulation of Candida albicans ABC transporters CDR1 and CDR2. Eukaryot Cell 3, 1639–1652.[CrossRef]
    [Google Scholar]
  8. Coste, A., Turner, V., Ischer, F., Morschhäuser, J., Forche, A., Semelcki, A., Berman, J., Bille, J. & Sanglard, D. ( 2006; ). A mutation in Tac1p, a transcription factor regulating CDR1 and CDR2, is coupled with loss of heterozygosity at chromosome 5 to mediate antifungal resistance in Candida albicans. Genetics 172, 2139–2156.
    [Google Scholar]
  9. Coste, A., Selmecki, A., Forche, A., Diogo, D., Bougnoux, M. E., d'Enfert, C., Berman, J. & Sanglard, D. ( 2007; ). Genotypic evolution of azole resistance mechanisms in sequential Candida albicans isolates. Eukaryot Cell 6, 1889–1904.[CrossRef]
    [Google Scholar]
  10. De Micheli, M., Bille, J., Schueller, C. & Sanglard, D. ( 2002; ). A common drug-responsive element mediates the upregulation of the Candida albicans ABC transporters CDR1 and CDR2, two genes involved in antifungal drug resistance. Mol Microbiol 43, 1197–1214.[CrossRef]
    [Google Scholar]
  11. Decottignies, A., Grant, A. M., Nichols, J. W., de Wet, H., McIntosh, D. B. & Goffeau, A. ( 1998; ). ATPase and multidrug transport activities of the overexpressed yeast ABC protein Yor1p. J Biol Chem 273, 12612–12622.[CrossRef]
    [Google Scholar]
  12. Delaveau, T., Delahodde, A., Carvajal, E., Subik, J. & Jacq, C. ( 1994; ). PDR3, a new yeast regulatory gene, is homologous to PDR1 and controls the multidrug resistance phenomenon. Mol Gen Genet 244, 501–511.[CrossRef]
    [Google Scholar]
  13. DeRisi, J., van den Hazel, B., Marc, P., Balzi, E., Brown, P., Jacq, C. & Goffeau, A. ( 2000; ). Genome microarray analysis of transcriptional activation in multidrug resistance yeast mutants. FEBS Lett 470, 156–160.[CrossRef]
    [Google Scholar]
  14. Devaux, F., Marc, P., Bouchoux, C., Delaveau, T., Hikkel, I., Potier, M. C. & Jacq, C. ( 2001; ). An artificial transcription activator mimics the genome-wide properties of the yeast Pdr1 transcription factor. EMBO Rep 2, 493–498.[CrossRef]
    [Google Scholar]
  15. Feinberg, A. P. & Vogelstein, B. ( 1984; ). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137, 266–267.[CrossRef]
    [Google Scholar]
  16. Fonzi, W. A. & Irwin, M. Y. ( 1993; ). Isogenic strain construction and gene mapping in Candida albicans. Genetics 134, 717–728.
    [Google Scholar]
  17. Gietz, D., St. Jean, A., Woods, R. A. & Schiestl, R. H. ( 1992; ). Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20, 1425 [CrossRef]
    [Google Scholar]
  18. Gupta, V., Kohli, A., Krishnamurthy, S., Puri, N., Aalamgeer, S. A., Panwar, S. & Prasad, R. ( 1998; ). Identification of polymorphic mutant alleles of CaMDR1, a major facilitator of Candida albicans which confers multidrug resistance, and its in vitro transcriptional activation. Curr Genet 34, 192–199.[CrossRef]
    [Google Scholar]
  19. Hanahan, D. ( 1985; ). Techniques for transformation of E. coli. In DNA Cloning: a Practical Approach. Edited by D. M. Glover. Oxford: IRL Press.
  20. Harry, J. B., Oliver, B. G., Song, J. L., Silver, P. M., Little, J. T., Choiniere, J. & White, T. C. ( 2005; ). Drug-induced regulation of the MDR1 promoter in Candida albicans. Antimicrob Agents Chemother 49, 2785–2792.[CrossRef]
    [Google Scholar]
  21. Hiller, D., Sanglard, D. & Morschhauser, J. ( 2006a; ). Overexpression of the MDR1 gene is sufficient to confer increased resistance to toxic compounds in Candida albicans. Antimicrob Agents Chemother 50, 1365–1371.[CrossRef]
    [Google Scholar]
  22. Hiller, D., Stahl, S. & Morschhauser, J. ( 2006b; ). Multiple cis-acting sequences mediate upregulation of the MDR1 efflux pump in a fluconazole-resistant clinical Candida albicans isolate. Antimicrob Agents Chemother 50, 2300–2308.[CrossRef]
    [Google Scholar]
  23. Kaiser, B., Munder, T., Saluz, H. P., Kunkel, W. & Eck, R. ( 1999; ). Identification of a gene encoding the pyruvate decarboxylase gene regulator CaPdc2p from Candida albicans. Yeast 15, 585–591.[CrossRef]
    [Google Scholar]
  24. Karababa, M., Coste, A. T., Rognon, B., Bille, J. & Sanglard, D. ( 2004; ). Comparison of gene expression profiles of Candida albicans azole-resistant clinical isolates and laboratory strains exposed to drugs inducing multidrug transporters. Antimicrob Agents Chemother 48, 3064–3079.[CrossRef]
    [Google Scholar]
  25. Karpichev, I. V. & Small, G. M. ( 1998; ). Global regulatory functions of Oaf1p and Pip2p (Oaf2p), transcription factors that regulate genes encoding peroxisomal proteins in Saccharomyces cerevisiae. Mol Cell Biol 18, 6560–6570.
    [Google Scholar]
  26. Katzmann, D. J., Hallstrom, T. C., Mahe, Y. & Moye-Rowley, W. S. ( 1996; ). Multiple Pdr1p/Pdr3p binding sites are essential for normal expression of the ATP binding cassette transporter protein-encoding gene PDR5. J Biol Chem 271, 23049–23054.[CrossRef]
    [Google Scholar]
  27. Leberer, E., Harcus, D., Broadbent, I. D., Clark, K. L., Dignard, D., Ziegelbauer, K., Schmidt, A., Gow, N. A., Brown, A. J. & Thomas, D. Y. ( 1996; ). Signal transduction through homologs of the Ste20p and Ste7p protein kinases can trigger hyphal formation in the pathogenic fungus Candida albicans. Proc Natl Acad Sci U S A 93, 13217–13222.[CrossRef]
    [Google Scholar]
  28. Longtine, M. S., McKenzie, A., III, Demarini, D. J., Shah, N. G., Wach, A., Brachat, A., Philippsen, P. & Pringle, J. R. ( 1998; ). Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14, 953–961.[CrossRef]
    [Google Scholar]
  29. MacPherson, S., Larochelle, M. & Turcotte, B. ( 2006; ). A fungal family of transcriptional regulators: the zinc cluster proteins. Microbiol Mol Biol Rev 70, 583–604.[CrossRef]
    [Google Scholar]
  30. Martchenko, M., Levitin, A., Hogues, H., Nantel, A. & Whiteway, M. ( 2007; ). Transcriptional rewiring of fungal galactose-metabolism circuitry. Curr Biol 17, 1007–1013.[CrossRef]
    [Google Scholar]
  31. Morschhauser, J., Barker, K. S., Liu, T. T., Bla, B. W. J., Homayouni, R. & Rogers, P. D. ( 2007; ). The transcription factor Mrr1p controls expression of the MDR1 efflux pump and mediates multidrug resistance in Candida albicans. PLoS Pathog 3, e164 [CrossRef]
    [Google Scholar]
  32. Nourani, A., Papajova, D., Delahodde, A., Jacq, C. & Subik, J. ( 1997; ). Clustered amino acid substitutions in the yeast transcription regulator Pdr3p increase pleiotropic drug resistance and identify a new central regulatory domain. Mol Gen Genet 256, 397–405.[CrossRef]
    [Google Scholar]
  33. Phillips, A. J., Sudbery, I. & Ramsdale, M. ( 2003; ). Apoptosis induced by environmental stresses and amphotericin B in Candida albicans. Proc Natl Acad Sci U S A 100, 14327–14332.[CrossRef]
    [Google Scholar]
  34. Phillips, A. J., Crowe, J. D. & Ramsdale, M. ( 2006; ). Ras pathway signaling accelerates programmed cell death in the pathogenic fungus Candida albicans. Proc Natl Acad Sci U S A 103, 726–731.[CrossRef]
    [Google Scholar]
  35. Riggle, P. J. & Kumamoto, C. A. ( 2006; ). Transcriptional regulation of MDR1, encoding a drug efflux determinant, in fluconazole-resistant Candida albicans strains through an Mcm1p binding site. Eukaryot Cell 5, 1957–1968.[CrossRef]
    [Google Scholar]
  36. Rognon, B., Kozovska, Z., Coste, A. T., Pardini, G. & Sanglard, D. ( 2006; ). Identification of promoter elements responsible for the regulation of MDR1 from Candida albicans, a major facilitator transporter involved in azole resistance. Microbiology 152, 3701–3722.[CrossRef]
    [Google Scholar]
  37. Sanglard, D., Kuchler, K., Ischer, F., Pagani, J. L., Monod, M. & Bille, J. ( 1995; ). Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters. Antimicrob Agents Chemother 39, 2378–2386.[CrossRef]
    [Google Scholar]
  38. Sanglard, D., Ischer, F., Monod, M. & Bille, J. ( 1996; ). Susceptibilities of Candida albicans multidrug transporter mutants to various antifungal agents and other metabolic inhibitors. Antimicrob Agents Chemother 40, 2300–2305.
    [Google Scholar]
  39. Sanglard, D., Ischer, F., Monod, M. & Bille, J. ( 1997; ). Cloning of Candida albicans genes conferring resistance to azole antifungal agents: characterization of CDR2, a new multidrug ABC-transporter gene. Microbiology 143, 405–416.[CrossRef]
    [Google Scholar]
  40. Shen, H., An, M. M., Wang de, J., Xu, Z., Zhang, J. D., Gao, P. H., Cao, Y. Y., Cao, Y. B. & Jiang, Y. Y. ( 2007; ). Fcr1p inhibits development of fluconazole resistance in Candida albicans by abolishing CDR1 induction. Biol Pharm Bull 30, 68–73.[CrossRef]
    [Google Scholar]
  41. Talibi, D. & Raymond, M. ( 1999; ). Isolation of a putative Candida albicans transcriptional regulator involved in pleiotropic drug resistance by functional complementation of a pdr1 pdr3 mutation in Saccharomyces cerevisiae. J Bacteriol 181, 231–240.
    [Google Scholar]
  42. Wolfger, H., Mahe, Y., Parle-McDermott, A., Delahodde, A. & Kuchler, K. ( 1997; ). The yeast ATP binding cassette (ABC) protein genes PDR10 and PDR15 are novel targets for the Pdr1 and Pdr3 transcriptional regulators. FEBS Lett 418, 269–274.[CrossRef]
    [Google Scholar]
  43. Yang, X., Talibi, D., Weber, S., Poisson, G. & Raymond, M. ( 2001; ). Functional isolation of the Candida albicans FCR3 gene encoding a bZip transcription factor homologous to Saccharomyces cerevisiae Yap3p. Yeast 18, 1217–1225.[CrossRef]
    [Google Scholar]
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