Open AccessA yeast cell‐based system for screening Candida glabrata multidrug resistance reversal agents and selection of loss‐of‐function pdr1 mutants
Author(s) -
Goffa Eduard,
Bialkova Alexandra,
Batova Monika,
Dzugasova Vladimira,
Subik Julius
Publication year - 2011
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2010.00702.x
Subject(s) - biology , efflux , mutant , multiple drug resistance , saccharomyces cerevisiae , candida glabrata , yeast , genetic screen , gene , atp binding cassette transporter , mutation , phenotype , drug resistance , transcription factor , function (biology) , genetics , transporter , candida albicans
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. Cg Pdr1p 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 Cg Pdr1p 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 Cg Pdr1p‐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 Cg Pdr1p activity or loss‐of‐function Cgpdr1 mutations, and is amenable to high‐throughput screening on solid or liquid media.