Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Biochemistry & Molecular Biology
A Novel Screening for Inhibitors of a Pleiotropic Drug Resistant Pump, Pdr5, in Saccharomyces cerevisiae
Kazumi HIRAGAAnoja WANIGASEKERAHisanori SUGINobuyuki HAMANAKAKohei ODA
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2001 Volume 65 Issue 7 Pages 1589-1595

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Abstract

Yeast is an excellent model system of eukaryotes for the study of molecular mechanisms of ATP-binding cassette transporters. Pdr5 protein is a yeast Saccharomyces cerevisiae ATP-binding cassette transporter conferring resistance to several unrelated drugs. Here, we described a novel drug screening system designated to detect compounds that inhibit the function of Pdr5. An indicator strain with increased drug sensitivity was constructed with an ergosterol-deficient background (Δsyr1/erg3 null mutation). The sensitivity of the indicator strain (Δsyr1/erg3Δpdr5Δsnq2) to the Pdr5 substrates, cycloheximide and cerulenin, was increased 16-fold and 4-fold against wild type, respectively. The screening system is mainly based on the growth inhibition of the PDR5-overexpressed indicator strain with the combination of a sample and cycloheximide or cerulenin. The effect of an mdr inhibitor, FK506 on the screening system was clearly detected even at a low concentration (∼0.5 μg/ml). In addition, accumulation of rhodamine 6G in the cells was detected as a result of Pdr5 inhibition by FK506. These results indicated that the screening system is useful for a sensitive screening of Pdr5-specific inhibitors with low toxicity.

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© 2001 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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