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A proteomic approach to understanding the development of multidrug-resistant Candida albicans strains

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Abstract

Resistance of the pathogenic yeast Candida albicans to the antifungal agent fluconazole is often caused by the overexpression of genes that encode multidrug efflux pumps (CDR1, CDR2, or MDR1). We have undertaken a proteomic approach to gain further insight into the regulatory network controlling efflux pump expression and drug resistance in C. albicans. Three pairs of matched fluconazole-susceptible and resistant clinical C. albicans isolates, in which drug resistance correlated with stable activation of MDR1 or CDR1/2, were analyzed for differences in their protein expression profiles. In two independent, MDR1-overexpressing, strains, additional up-regulated proteins were identified, which are encoded by the YPR127 gene and several members of the IFD (YPL088) gene family. All are putative aldo-keto reductases of unknown function. These proteins were not up-regulated in a fluconazole-resistant strain that overexpressed CDR1 and CDR2 but not MDR1, indicating that expression of the various efflux pumps of C. albicans is controlled by different regulatory networks. To investigate the possible role of YPR127 in the resistance phenotype of the clinical isolates, we constitutively overexpressed the gene in a C. albicans laboratory strain. In addition, the gene was deleted in a C. albicans laboratory strain and in one of the drug-resistant clinical isolates in which it was overexpressed. Neither forced overexpression nor deletion of YPR127 affected the susceptibility of the strains to drugs and other toxic substances, suggesting that the regulatory networks which control the expression of efflux pumps in C. albicans also control genes involved in cellular functions not related to drug resistance.

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Acknowledgements

This study was supported by grants from the Deutsche Forschungsgemeinschaft (DFG grant MO846/3), the European Community (EC project QLK2-CT-2001–02377) and the Bayerische Forschungsstiftung. Joachim Morschhäuser was the recipient of a Heisenberg fellowship from the Deutsche Forschungsgemeinschaft. We thank Dörte Becher and Dirk Albrecht for support in protein identification. Sequence data for Candida albicans was obtained from the Stanford Genome Technology Center website (http://www-sequence.stanford.edu/group/candida). Sequencing of Candida albicans was accomplished with the support of the NIDR and the Burroughs Wellcome Fund. The work has been carried out in compliance with the current laws governing genetic experimentation in Germany.

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Authors and Affiliations

  1. Institut für Molekulare Infektionsbiologie, Julius-Maximilians-Universität Würzburg, Röntgenring 11, 97070, Würzburg, Germany

    H. Kusch, K. Biswas, S. Schwanfelder & J. Morschhäuser

  2. Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, 17489, Greifswald, Germany

    H. Kusch, S. Engelmann & M. Hecker

  3. Departments of Clinical Pharmacy and Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, 26 South Dunlap, Suite 513, Memphis, TN 38163, USA

    P. D. Rogers

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  1. H. Kusch
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  2. K. Biswas
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  3. S. Schwanfelder
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  4. S. Engelmann
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  5. P. D. Rogers
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  6. M. Hecker
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  7. J. Morschhäuser
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Correspondence to J. Morschhäuser.

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Communicated by D. Y. Thomas

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Kusch, H., Biswas, K., Schwanfelder, S. et al. A proteomic approach to understanding the development of multidrug-resistant Candida albicans strains. Mol Genet Genomics 271, 554–565 (2004). https://doi.org/10.1007/s00438-004-0984-x

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  • DOI: https://doi.org/10.1007/s00438-004-0984-x

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