1887

Microbiology Society logo

Volume 156, Issue 8

Overexpression of restores respiratory growth and fluconazole sensitivity to a Δ mutant Free

Abstract

The yeast-like fungus favours respiration as a mechanism of energy production, and thus depends heavily on mitochondrial function. Previous studies of a Δ mutant revealed reduced expression of the mitochondrial elongation factor and defects in glycerol utilization, consistent with mitochondrial dysfunction. In this study, we found that expression of in the Δ mutant suppressed the mitochondrial defects, including growth on respiration-dependent carbon sources and fluconazole resistance, associated with deletion. Tetracycline, an inhibitor of mitochondrial translation, was found to confer resistance to fluconazole in the wild-type and Δ mutant, whereas the fluconazole susceptibility of the -overexpressing strain was unaffected by tetracycline treatment. In the presence of fluconazole, the Δ mutant exhibited increased activation of the global transcriptional regulator Sre1. overexpression failed to alter cleavage of Sre1 in response to fluconazole in the Δ mutant, suggesting that repression in the Δ mutant is distal to Sre1, or that it occurs through an independent pathway.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): qRT-PCR, quantitative real-time PCR
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.035923-0
2010-08-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/156/8/2558.html?itemId=/content/journal/micro/10.1099/mic.0.035923-0&mimeType=html&fmt=ahah

References

  1. Akhter S., McDade H. C., Gorlach J. M., Heinrich G., Cox G. M., Perfect J. R. 2003; Role of alternative oxidase gene in pathogenesis of Cryptococcus neoformans. Infect Immun 71:5794–5802
     [Google Scholar]
  2. Arthington-Skaggs B. A., Jradi H., Desai T., Morrison C. J. 1999; Quantitation of ergosterol content: novel method for determination of fluconazole susceptibility of Candida albicans. J Clin Microbiol 37:3332–3337
     [Google Scholar]
  3. Bicanic T., Harrison T. S. 2005; Cryptococcal meningitis. Br Med Bull 72:99–118
     [Google Scholar]
  4. Bicanic T., Harrison T., Niepieklo A., Dyakopu N., Meintjes G. 2006; Symptomatic relapse of HIV-associated cryptococcal meningitis after initial fluconazole monotherapy: the role of fluconazole resistance and immune reconstitution. Clin Infect Dis 43:1069–1073
     [Google Scholar]
  5. Brun S., Berges T., Poupard P., Vauzelle-Moreau C., Renier G., Chabasse D., Bouchara J. P. 2004; Mechanisms of azole resistance in petite mutants of Candida glabrata. Antimicrob Agents Chemother 48:1788–1796
     [Google Scholar]
  6. Chang Y. C., Bien C. M., Lee H., Espenshade P. J., Kwon-Chung K. J. 2007; Sre1p, a regulator of oxygen sensing and sterol homeostasis, is required for virulence in Cryptococcus neoformans. Mol Microbiol 64:614–629
     [Google Scholar]
  7. Chiron S., Suleau A., Bonnefoy N. 2005; Mitochondrial translation: elongation factor Tu is essential in fission yeast and depends on an exchange factor conserved in humans but not in budding yeast. Genetics 169:1891–1901
     [Google Scholar]
  8. Chun C. D., Liu O. W., Madhani H. D. 2007; A link between virulence and homeostatic responses to hypoxia during infection by the human fungal pathogen Cryptococcus neoformans. PLoS Pathog 3:e22
     [Google Scholar]
  9. De Luca C., Besagni C., Frontali L., Bolotin-Fukuhara M., Francisci S. 2006; Mutations in yeast mt tRNAs: specific and general suppression by nuclear encoded tRNA interactors. Gene 377:169–176
     [Google Scholar]
  10. Hughes A. L., Lee C. Y., Bien C. M., Espenshade P. J. 2007; 4-Methyl sterols regulate fission yeast SREBP-Scap under low oxygen and cell stress. J Biol Chem 282:24388–24396
     [Google Scholar]
  11. Husain S., Wagener M. M., Singh N. 2001; Cryptococcus neoformans infection in organ transplant recipients: variables influencing clinical characteristics and outcome. Emerg Infect Dis 7:375–381
     [Google Scholar]
  12. Ingavale S. S., Chang Y. C., Lee H., McClelland C. M., Leong M. L., Kwon-Chung K. J. 2008; Importance of mitochondria in survival of Cryptococcus neoformans under low oxygen conditions and tolerance to cobalt chloride. PLoS Pathog 4:e1000155
     [Google Scholar]
  13. Litter J., Keszthelyi A., Hamari Z., Pfeiffer I., Kucsera J. 2005; Differences in mitochondrial genome organization of Cryptococcus neoformans strains. Antonie Van Leeuwenhoek 88:249–255
     [Google Scholar]
  14. Liu X., Hu G., Panepinto J., Williamson P. R. 2006; Role of a VPS41 homologue in starvation response, intracellular survival and virulence of Cryptococcus neoformans. Mol Microbiol 61:1132–1146
     [Google Scholar]
  15. Livak K. J., Schmittgen T. D. 2001; Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔ T method. Methods 25:402–408
     [Google Scholar]
  16. Oliver B. G., Silver P. M., Marie C., Hoot S. J., Leyde S. E., White T. C. 2008; Tetracycline alters drug susceptibility in Candida albicans and other pathogenic fungi. Microbiology 154:960–970
     [Google Scholar]
  17. Panepinto J., Liu L., Ramos J., Zhu X., Valyi-Nagy T., Eksi S., Fu J., Jaffe H. A., Wickes B., Williamson P. R. 2005; The DEAD-box RNA helicase Vad1 regulates multiple virulence-associated genes in Cryptococcus neoformans. J Clin Invest 115:632–641
     [Google Scholar]
  18. Panepinto J., Komperda K. W., Frases S., Park Y. D., Djordejevic J. T., Casadevall A., Williamson P. R. 2009; Sec6-dependent sorting of fungal extracellular exosomes and laccase of Cryptococcus neoformans. Mol Microbiol 71:1165–1176
     [Google Scholar]
  19. Perfect J. R., Durack D. T. 1985; Penetration of imidazoles and triazoles into cerebrospinal fluid of rabbits. J Antimicrob Chemother 16:81–86
     [Google Scholar]
  20. Petrou M. A., Shanson D. C. 2000; Susceptibility of Cryptococcus neoformans by the NCCLS microdilution and Etest methods using five defined media. J Antimicrob Chemother 46:815–818
     [Google Scholar]
  21. Rosenthal L. P., Bodley J. W. 1987; Purification and characterization of Saccharomyces cerevisiae mitochondrial elongation factor Tu. J Biol Chem 262:10955–10959
     [Google Scholar]
  22. Singh N., Lortholary O., Alexander B. D., Gupta K. L., John G. T., Pursell K., Munoz P., Klintmalm G. B., Stosor V. other authors 2005; Allograft loss in renal transplant recipients with Cryptococcus neoformans associated immune reconstitution syndrome. Transplantation 80:1131–1133
     [Google Scholar]
  23. Zhang L., Ging N. C., Komoda T., Hanada T., Suzuki T., Watanabe K. 2005; Antibiotic susceptibility of mammalian mitochondrial translation. FEBS Lett 579:6423–6427
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.035923-0
Loading
Overexpression of TUF1 restores respiratory growth and fluconazole sensitivity to a Cryptococcus neoformansvad1Δ mutant
Microbiology 156, 2558 (2010); https://doi.org/10.1099/mic.0.035923-0
/content/journal/micro/10.1099/mic.0.035923-0
/content/journal/micro/10.1099/mic.0.035923-0
Loading

Data & Media loading...

Most cited 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