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Originally published In Press as doi:10.1074/jbc.M208549200 on January 14, 2003

J. Biol. Chem., Vol. 278, Issue 13, 11427-11432, March 28, 2003
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A General Strategy to Uncover Transcription Factor Properties Identifies a New Regulator of Drug Resistance in Yeast*

Imrich HikkelDagger §, Ancuta Lucau-DanilaDagger , Thierry Delaveau, Philippe Marc, Frédéric Devaux, and Claude Jacq||

From the Laboratoire de Génétique Moléculaire, CNRS UMR8541, Ecole Normale Supérieure, 46 rue d'Ulm 75230 Paris Cedex 05, France

We demonstrate a genomewide approach to determine the physiological role of a putative transcription factor, Ylr266, identified through yeast genome sequencing program. We constructed activated forms of the zinc finger (Zn2Cys6) protein Ylr266, and we analyzed the corresponding transcriptomes with DNA microarrays to characterize the up-regulated genes. The direct target genes of Ylr266 were further identified by in vivo chromatin immunoprecipitation procedure. The functions of the genes directly controlled by YLR266c are in agreement with the observed drug-resistance phenotype of the cell expressing an activated form of Ylr266. These target genes code for ATP-binding cassette or major facilitator superfamily transporters such as PDR15, YOR1, or AZR1 or for other proteins such as SNG1, YJL216c, or YLL056c which are already known to be involved in the yeast pleiotropic drug resistance (PDR) phenomenon. YLR266c could thus be named PDR8. Overlaps with the other PDR networks argue in favor of a new specific role for PDR8 in connection with the well known PDR regulators PDR1/PDR3 and YRR1. This strategy to identify the regulatory properties of an anonymous transcription factor is likely to be generalized to all the Zn2Cys6 transcription factors from Saccharomyces cerevisiae and related yeasts.

* This work was supported in part by grants from CNRS (Puces a ADN 2A2112) and Association pour la Recherche contre le Cancer (5691).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Both authors contributed equally to this work.

§ Present address: Comenius University, Dept. of Microbiology and Virology, Faculty of Natural Sciences, Mlynska Dolina B2, 842 15 Bratislava, Slovak Republic.

Supported by a European Commission long-term fellowship (Combating of MDR, QLK2-CT.2001.02377).

|| To whom correspondence should be addressed. E-mail: jacq@biologie.ens.fr.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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