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Gene expression and proteomic profile analysis reveals novel gene and protein expression patterns associated with fluconazole resistance in clinical isolates of Candida albicans


, : Gene expression and proteomic profile analysis reveals novel gene and protein expression patterns associated with fluconazole resistance in clinical isolates of Candida albicans. Abstracts of the Interscience Conference on Antimicrobial Agents & Chemotherapy 43: 442

Background: In order to identify novel mechanisms of azole resistance, we examined changes in the gene expression and proteomic profiles of two series of C. albicans isolates representing the acquisition of fluconazole resistance. These sets were chosen as their acquisition of resistance is independent of over-expression of the efflux pump genes CDR1, CDR2, or MDR1. Methods: Isolates were from an AIDS patient with oropharyngeal candidiasis who failed fluconazole therapy (isolates S1, MIC=1 mug/mL and S2, MIC>64 mug/mL; M1, MIC=1 and M4, MIC=32 mug/ml). Gene expression profiles were compared between isolates S1 and S2 as well as M1 and M4 in replicate experiments. Genes of interest were verified for differential expression by RT-PCR. The protein fraction was subjected to 2D-SDS-PAGE. Protein spots were identified by MALDI-TOF-TOF MS peptide mass fingerprinting using a C. albicans ORF database formatted for PROWL software. Results: We found 12 genes to be up-regulated and 34 genes to be down-regulated in set S, and 12 genes to be up-regulated and 26 genes to be down-regulated in set M. Of these, 22 were similarly differentially expressed in both sets. Up-regulated genes included CDC31, KAP95, MRPL23A, IPF3607, IPF3950, and IPF7260. Down-regulated genes included CSE1, ERG24, HSP60, LYS12, SEC31, IPF11153, IPF19862, IPF29, IPF5533, IPF6459, IPF6600, IPF7204, IPF8374, IPF8810, and IPF883. Differential expression of CDR1, CDR2, MDR1, and ERG11 was not observed. Nine proteins were found to be differentially expressed in set S and 15 in set M. Differentially expressed proteins in set M included the down-regulation of Erg10p and Hem13p. Conclusions: These results suggest novel mechanisms of azole resistance. Further evaluation of the role of these genes and proteins in this process is warranted.

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