RRC ID 46775
Author Pais P, Costa C, Pires C, Shimizu K, Chibana H, Teixeira MC.
Title Membrane Proteome-Wide Response to the Antifungal Drug Clotrimazole in Candida glabrata: Role of the Transcription Factor CgPdr1 and the Drug:H+ Antiporters CgTpo1_1 and CgTpo1_2.
Journal Mol Cell Proteomics
Abstract Azoles are widely used antifungal drugs. This family of compounds includes triazoles, mostly used in the treatment of systemic infections, and imidazoles, such as clotrimazole, often used in the case of superficial infections. Candida glabrata is the second most common cause of candidemia worldwide and presents higher levels of intrinsic azole resistance when compared with Candida albicans, thus being an interesting subject for the study of azole resistance mechanisms in fungal pathogens.Since resistance often relies on the action of membrane transporters, including drug efflux pumps from the ATP-binding cassette family or from the Drug:H(+) antiporter (DHA)(1) family, an iTRAQ-based membrane proteomics analysis was performed to identify all the membrane-associated proteins whose abundance changes in C. glabrata cells exposed to the azole drug clotrimazole. Proteins found to have significant expression changes in this context were clustered into functional groups, namely: glucose metabolism, oxidative phosphorylation, mitochondrial import, ribosome components and translation machinery, lipid metabolism, multidrug resistance transporters, cell wall assembly, and stress response, comprising a total of 37 proteins. Among these, the DHA transporter CgTpo1_2 (ORF CAGL0E03674g) was identified as overexpressed in the C. glabrata membrane in response to clotrimazole. Functional characterization of this putative drug:H(+) antiporter, and of its homolog CgTpo1_1 (ORF CAGL0G03927g), allowed the identification of these proteins as localized to the plasma membrane and conferring azole drug resistance in this fungal pathogen by actively extruding the drug to the external medium. The cell wall protein CgGas1 was also shown to confer azole drug resistance through cell wall remodeling. Finally, the transcription factor CgPdr1 in the clotrimazole response was observed to control the expression of 20 of the identified proteins, thus highlighting the existence of additional unforeseen targets of this transcription factor, recognized as a major regulator of azole drug resistance in clinical isolates.
Volume 15(1)
Pages 57-72
Published 2016-1-1
DOI 10.1074/mcp.M114.045344
PII S1535-9476(20)33708-7
PMID 26512119
PMC PMC4762512
MeSH Antifungal Agents / pharmacology Antiporters / genetics Antiporters / metabolism Candida glabrata / drug effects* Candida glabrata / genetics Candida glabrata / metabolism Cell Membrane / drug effects Cell Membrane / metabolism Clotrimazole / pharmacology* Drug Resistance, Fungal / drug effects Drug Resistance, Fungal / genetics Fungal Proteins / genetics Fungal Proteins / metabolism* Gene Expression Regulation, Fungal Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Membrane Proteins / genetics Membrane Proteins / metabolism* Microscopy, Fluorescence Mutation Proteome / genetics Proteome / metabolism* Proteomics / methods* Reverse Transcriptase Polymerase Chain Reaction Transcription Factors / genetics Transcription Factors / metabolism
IF 4.87
Times Cited 18
Pathogenic microorganisms NA