1887

Microbiology

Volume 146, Issue 11

A novel multidrug efflux transporter gene of the major facilitator superfamily from () conferring resistance to fluconazole Free

Abstract

The GenBank accession number for the sequence reported in this paper is AF188621.

Azole resistance in can be mediated by several resistance mechanisms. Among these, alterations of the azole target enzyme and the overexpression of multidrug efflux transporter genes are the most frequent. To identify additional putative azole resistance genes in , a genomic library from this organism was screened for complementation of fluconazole hypersusceptibility in YKKB-13 lacking the ABC (TP-inding assette) transporter gene . Among the genes obtained, a new gene was isolated and named (conazole resistance). The deduced amino acid sequence of showed similarity to (formerly ), a member of the major facilitator superfamily of multidrug efflux transporters. The expression of in YKKB-13 mediated not only resistance to fluconazole but also to cycloheximide among the different drugs tested. The disruption of in had only a slight effect on fluconazole susceptibility; however, it resulted in hypersusceptibility to mycophenolic acid, thus suggesting that this compound could be a substrate for the protein encoded by . Disruption of in a background of mutants with deletions in several multidrug efflux transporter genes, including , and , resulted in enhanced susceptibility to several azole derivatives. expression did not vary significantly between several pairs of azole-susceptible and azole-resistant clinical isolates. Therefore, seems not to be required for the development of azole resistance in clinical isolates.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): azole antifungal agents , Candida albicans , MF, major facilitator , MFS, major facilitator superfamily , multidrug efflux transporters and OPC, oropharyngeal candidiasis
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-11-2743
2000-11-01
2019-08-19

  • From This Site

    /content/journal/micro/10.1099/00221287-146-11-2743
    dcterms_title,dcterms_subject,pub_keyword
    10
    5
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/11/1462743a.html?itemId=/content/journal/micro/10.1099/00221287-146-11-2743&mimeType=html&fmt=ahah

References

  1. Alarco, A. M., Balan, I., Talibi, D., Mainville, N. & Raymond, M. ( 1997; ). AP1-mediated multidrug resistance in Saccharomyces cerevisiae requires FLR1 encoding a transporter of the major facilitator superfamily. J Biol Chem 272, 19304-19313.[CrossRef]
    [Google Scholar]
  2. Amakasu, H., Suzuki, Y., Nishizawa, M. & Fukasawa, T. ( 1993; ). Isolation and characterization of SGE1: a yeast gene that partially suppresses the gal11 mutation in multiple copies. Genetics 134, 675-683.
     [Google Scholar]
  3. Ben-Yaacov, R., Knoller, S., Caldwell, G. A., Becker, J. M. & Koltin, Y. ( 1994; ). Candida albicans gene encoding resistance to benomyl and methotrexate is a multidrug resistance gene. Antimicrob Agents Chemother 38, 648-652.[CrossRef]
    [Google Scholar]
  4. Bissinger, P. H. & Kuchler, K. ( 1994; ). Molecular cloning and expression of the Saccharomyces cerevisiae STS1 gene product. A yeast ABC transporter conferring mycotoxin resistance. J Biol Chem 269, 4180-4186.
     [Google Scholar]
  5. Boerlin, P., Boerlin-Petzold, F., Goudet, J., Durussel, C., Pagani, J. L., Chave, J. P. & Bille, J. ( 1996; ). Typing Candida albicans oral isolates from human immunodeficiency virus-infected patients by multilocus enzyme electrophoresis and DNA fingerprinting. J Clin Microbiol 34, 1235-1248.
     [Google Scholar]
  6. Chen, W. & Struhl, K. ( 1988; ). Saturation mutagenesis of a yeast his3 ‘TATA element’: genetic evidence for a specific TATA-binding protein. Proc Natl Acad Sci USA 85, 2691-2695.[CrossRef]
    [Google Scholar]
  7. Chibana, H., Magee, B. B., Grindle, S., Ran, Y., Scherer, S. & Magee, P. T. ( 1998; ). A physical map of chromosome 7 of Candida albicans. Genetics 149, 1739-1752.
     [Google Scholar]
  8. Decottignies, A., Lambert, L., Catty, P., Degand, H., Epping, E. A., Moye-Rowley, W. S., Balzi, E. & Goffeau, A. ( 1995; ). Identification and characterization of SNQ2, a new multidrug ATP binding cassette transporter of the yeast plasma membrane. J Biol Chem 270, 18150-18157.[CrossRef]
    [Google Scholar]
  9. Delling, U., Raymond, M. & Schurr, E. ( 1998; ). Identification of Saccharomyces cerevisiae genes conferring resistance to quinoline ring-containing antimalarial drugs. Antimicrob Agents Chemother 42, 1034-1041.
     [Google Scholar]
  10. Ehrenhofer-Murray, A. E., Wurgler, F. E. & Sengstag, C. ( 1994; ). The Saccharomyces cerevisiae SGE1 gene product: a novel drug-resistance protein within the major facilitator superfamily. Mol Gen Genet 244, 287-294.
     [Google Scholar]
  11. Ellenberger, T. E., Brandl, C. J., Struhl, K. & Harrison, S. C. ( 1992; ). The GCN4 basic region leucine zipper binds DNA as a dimer of uninterrupted alpha helices: crystal structure of the protein–DNA complex. Cell 71, 1223-1237.[CrossRef]
    [Google Scholar]
  12. 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]
  13. Fling, M. E., Kopf, J., Tamarkin, A., Gorman, J. A., Smith, H. A. & Koltin, Y. ( 1991; ). Analysis of a Candida albicans gene that encodes a novel mechanism for resistance to benomyl and methotrexate. Mol Gen Genet 227, 318-329.
     [Google Scholar]
  14. Fonzi, W. A. & Irwin, M. Y. ( 1993; ). Isogenic strain construction and gene mapping in Candida albicans. Genetics 134, 717-728.
     [Google Scholar]
  15. Goffeau, A., Park, J., Paulsen, I. T., Jonniaux, J. L., Dinh, T., Mordant, P. & Saier, M. H.Jr ( 1997; ). Multidrug-resistant transport proteins in yeast: complete inventory and phylogenetic characterization of yeast open reading frames with the major facilitator superfamily. Yeast 13, 43-54.[CrossRef]
    [Google Scholar]
  16. Goldway, M., Teff, D., Schmidt, R., Oppenheim, A. B. & Koltin, Y. ( 1995; ). Multidrug resistance in Candida albicans: disruption of the BEN r gene. Antimicrob Agents Chemother 39, 422-426.[CrossRef]
    [Google Scholar]
  17. Gompel-Klein, P. & Brendel, M. ( 1990; ). Allelism of SNQ1 and ATR1, genes of the yeast Saccharomyces cerevisiae required for controlling sensitivity to 4-nitroquinoline-N-oxide and aminotriazole. Curr Genet 18, 93-96.[CrossRef]
    [Google Scholar]
  18. Graham, T. R., Scott, P. A. & Emr, S. D. ( 1993; ). Brefeldin A reversibly blocks early but not late protein transport steps in the yeast secretory pathway. EMBO J 12, 869-877.
     [Google Scholar]
  19. Hanahan, D. ( 1985; ). Techniques for transformation of E. coli. In DNA Cloning. A Practical Approach, pp. 109-135. Edited by D. M. Glover. Oxford: IRL Press.
  20. Hube, B., Monod, M., Schofield, D. A., Brown, A. J. & Gow, N. A. ( 1994; ). Expression of seven members of the gene family encoding secretory aspartyl proteinases in Candida albicans. Mol Microbiol 14, 87-99.[CrossRef]
    [Google Scholar]
  21. Jia, Z. P., McCullough, N., Wong, L. & Young, P. G. ( 1993; ). The amiloride resistance gene, car1, of Schizosaccharomyces pombe. Mol Gen Genet 241, 298-304.
     [Google Scholar]
  22. Kanazawa, S., Driscoll, M. & Struhl, K. ( 1988; ). ATR1, a Saccharomyces cerevisiae gene encoding a transmembrane protein required for aminotriazole resistance. Mol Cell Biol 8, 664-673.
     [Google Scholar]
  23. Kim, J. & Struhl, K. ( 1995; ). Determinants of half-site spacing preferences that distinguish AP-1 and ATF/CREB bZIP domains. Nucleic Acids Res 23, 2531-2537.[CrossRef]
    [Google Scholar]
  24. Launhardt, H., Hinnen, A. & Munder, T. ( 1998; ). Drug-induced phenotypes provide a tool for the functional analysis of yeast genes. Yeast 14, 935-942.[CrossRef]
    [Google Scholar]
  25. Marichal, P. & Vanden Bossche, H. ( 1995; ). Mechanisms of resistance to azole antifungals. Acta Biochim Pol 42, 509-516.
     [Google Scholar]
  26. Morisaki, N., Funabashi, H., Shimazawa, R., Furukawa, J., Kawaguchi, A., Okuda, S. & Iwasaki, S. ( 1993; ). Effect of side-chain structure on inhibition of yeast fatty-acid synthase by cerulenin analogues. Eur J Biochem 211, 111-115.[CrossRef]
    [Google Scholar]
  27. Mutoh, E., Mochizuki, M., Ohta, A. & Takagi, M. ( 1995; ). Inducible expression of a gene encoding an L41 ribosomal protein responsible for the cycloheximide-resistant phenotype in the yeast Candida maltosa. J Bacteriol 177, 5383-5386.
     [Google Scholar]
  28. Nelissen, B., Mordant, P., Jonniaux, J. L., De Wachter, R. & Goffeau, A. ( 1995; ). Phylogenetic classification of the major superfamily of membrane transport facilitators, as deduced from yeast genome sequencing. FEBS Lett 377, 232-236.[CrossRef]
    [Google Scholar]
  29. Odds, F. C., Schmid, J. & Soll, D. R. ( 1990; ). Epidemiology of Candida infections in AIDS. In Mycoses in AIDS Patients, pp. 67-74. Edited by N. Y. P. Press. New York: Plenum.
  30. Odds, F. C., Dams, G., Just, G. & Lewi, P. ( 1996; ). Susceptibilities of Candida spp. to antifungal agents visualized by two-dimensional scatterplots of relative growth. Antimicrob Agents Chemother 40, 588-594.
     [Google Scholar]
  31. Oliphant, A. R., Brandl, C. J. & Struhl, K. ( 1989; ). Defining the sequence specificity of DNA-binding proteins by selecting binding sites from random-sequence oligonucleotides: analysis of yeast GCN4 protein. Mol Cell Biol 9, 2944-2949.
     [Google Scholar]
  32. Oskouian, B. & Saba, J. D. ( 1999; ). YAP1 confers resistance to the fatty acid synthase inhibitor cerulenin through the transporter Flr1p in Saccharomyces cerevisiae. Mol Gen Genet 261, 346-353.[CrossRef]
    [Google Scholar]
  33. Powderly, W. G. ( 1994; ). Resistant candidiasis. AIDS Res Hum Retrovir 10, 925-929.[CrossRef]
    [Google Scholar]
  34. Prasad, R., De Wergifosse, P., Goffeau, A. & Balzi, E. ( 1995; ). Molecular cloning and characterization of a novel gene of Candida albicans, CDR1, conferring multiple resistance to drugs and antifungals. Curr Genet 27, 320-329.[CrossRef]
    [Google Scholar]
  35. Rex, J. H., Pfaller, M. A., Barry, A. L., Nelson, P. W. & Webb, C. D. ( 1995; ). Antifungal susceptibility testing of isolates from a randomized, multicenter trial of fluconazole versus amphotericin B as treatment of nonneutropenic patients with candidemia. NIAID Mycoses Study Group and the Candidemia Study Group. Antimicrob Agents Chemother 39, 40-44.[CrossRef]
    [Google Scholar]
  36. 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]
  37. 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]
  38. 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]
  39. Sanglard, D., Ischer, F., Koymans, L. & Bille, J. ( 1998; ). Amino acid substitutions in the cytochrome P-450 lanosterol 14alpha-demethylase (CYP51A1) from azole-resistant Candida albicans clinical isolates contribute to resistance to azole antifungal agents. Antimicrob Agents Chemother 42, 241-253.[CrossRef]
    [Google Scholar]
  40. Sanglard, D., Ischer, F., Calabrese, D., Majcherczyk, P. A. & Bille, J. ( 1999; ). The ATP Binding Cassette (ABC)-transporter gene CgCDR1 from Candida glabrata is involved in the resistance of clinical isolates to azole antifungal agents. Antimicrob Agents Chemother 43, 2753-2765.
     [Google Scholar]
  41. Struhl, K., Chen, W., Hill, D. E., Hope, I. A. & Oettinger, M. A. ( 1985; ). Constitutive and coordinately regulated transcription of yeast genes: promoter elements, positive and negative regulatory sites, and DNA binding proteins. Cold Spring Harbor Symp Quant Biol 50, 489-503.[CrossRef]
    [Google Scholar]
  42. Troillet, N., Durussel, C., Bille, J., Glauser, M. P. & Chave, J. P. ( 1993; ). Correlation between in vitro susceptibility of Candida albicans and fluconazole-resistant oropharyngeal candidiasis in HIV-infected patients. Eur J Clin Microbiol Infect Dis 12, 911-915.[CrossRef]
    [Google Scholar]
  43. Vanden Bossche, H., Marichal, P. & Odds, F. C. ( 1994a; ). Molecular mechanisms of drug resistance in fungi. Trends Microbiol 2, 393-400.[CrossRef]
    [Google Scholar]
  44. Vanden Bossche, H., Warnock, D. W., Dupont, B. & 7 other authors ( 1994; b). Mechanisms and clinical impact of antifungal drug resistance. J Med Vet Mycol 32, 189–202.
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-146-11-2743
Loading
A novel multidrug efflux transporter gene of the major facilitator superfamily from Candida albicans (FLU1) conferring resistance to fluconazole
Microbiology 146, 2743 (2000); https://doi.org/10.1099/00221287-146-11-2743
/content/journal/micro/10.1099/00221287-146-11-2743
/content/journal/micro/10.1099/00221287-146-11-2743
Loading

Data & Media loading...

Most Cited This Month

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