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Updated: Jun 6, 2026
Rapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae
Published on: April 5, 2015
A yeast cell-based system for screening Candida glabrata multidrug resistance reversal agents and selection of
Eduard Goffa1, Alexandra Bialkova, Monika Batova
1Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovak Republic.
Abstract:
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.
Insights
Researchers developed a new screening system to identify compounds that inhibit the CgPdr1p transcription factor, a key driver of multidrug resistance in Candida glabrata yeast. This system aids in finding agents to reverse drug resistance.
Area of Science:
- Microbiology
- Molecular Biology
- Drug Discovery
Background:
- Multidrug resistance (MDR) in pathogenic yeast Candida glabrata is often linked to the overexpression of drug efflux pumps.
- This overexpression is frequently caused by gain-of-function mutations in the CgPDR1 gene, which encodes the CgPdr1p transcription factor.
- CgPdr1p regulates the expression of genes responsible for drug transport, making it a potential target for MDR reversal strategies.
Purpose of the Study:
- To establish a novel cell-based screening system for identifying compounds that inhibit the activity of the CgPdr1p transcription factor.
- To facilitate the discovery of agents that can sensitize multidrug-resistant yeast cells.
Main Methods:
- A heterologous screening system was developed using a hypersensitive Saccharomyces cerevisiae mutant strain.
- The system relies on the ability of compounds to rescue a growth defect caused by galactose-induced, CgPdr1p-driven overexpression of a dominant lethal pma1(D378N) allele.
- This lethal allele's expression is controlled by the ScPDR5 promoter.
Main Results:
- The developed system enables the rapid identification of compounds that inhibit CgPdr1p activity.
- It can also detect loss-of-function mutations in Cgpdr1.
- The screening method is adaptable for high-throughput screening on both solid and liquid media.
Conclusions:
- A robust and efficient cell-based screening system has been established for targeting CgPdr1p in Candida glabrata.
- This system is valuable for discovering novel multidrug resistance reversal agents.
- The approach holds promise for high-throughput screening in antifungal drug discovery.
