yeast cell-based system for screening Candida glabrata multidrug resistance reversal agents and selection of loss-of-function pdr1 mutants

In the pathogenic yeast Candida glabrata, multidrug resistance is associated with the overexpression of drug efflux pumps caused by gain-of-function mutations in the CgPDR1 gene. CgPdr1p transcription factor, which activates the expression of several drug efflux transporter genes, is considered to b...

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Bibliographic Details
Published in:FEMS yeast research 2011-03, Vol.11 (2), p.155-159
Main Authors: Goffa, Eduard, Bialkova, Alexandra, Batova, Monika, Dzugasova, Vladimira, Subik, Julius
Format: Article
Language:English
Subjects:
Candida glabrata
Candida glabrata - drug effects
Candida glabrata - genetics
CgPDR1
Cgpdr1 mutants
CgPdr1p inhibitors
Drug Evaluation, Preclinical - methods
Drug Resistance, Multiple, Fungal - drug effects
Fungal Proteins - genetics
Fungal Proteins - metabolism
Galactose
Gene Deletion
High-throughput screening
High-Throughput Screening Assays
Membrane Transport Proteins - genetics
Membrane Transport Proteins - metabolism
Microbial Viability - drug effects
Multidrug resistance
Multidrug resistant organisms
Mutants
Mutation
Proton-Translocating ATPases - genetics
Proton-Translocating ATPases - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae - drug effects
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
transcription factor
Yeast
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Summary:In the pathogenic yeast Candida glabrata, multidrug resistance is associated with the overexpression of drug efflux pumps caused by gain-of-function mutations in the CgPDR1 gene. CgPdr1p transcription factor, which activates the expression of several drug efflux transporter genes, is considered to be a promising target for compounds sensitizing the multidrug-resistant yeast cells. Here, we describe a cell-based screening system for detecting the inhibitory activity of compounds interfering with the CgPdr1p function in a heterologous genetic background of the hypersensitive Saccharomyces cerevisiae mutant strain. The screening is based on the ability to abrogate the growth defect of cells suffering from the galactose-induced and CgPdr1p-driven overexpression of a dominant lethal pma1(D378N) allele placed under the control of the ScPDR5 promoter. The system allows rapid identification of multidrug resistance reversal agents inhibiting the CgPdr1p activity or loss-of-function Cgpdr1 mutations, and is amenable to high-throughput screening on solid or liquid media.
ISSN:1567-1356
1567-1364
DOI:10.1111/j.1567-1364.2010.00702.x