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J. Biol. Chem., Vol. 278, Issue 30, 27605-27611, July 25, 2003

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Saccharomyces cerevisiae PIP2 Mediating Oleic Acid Induction and Peroxisome Proliferation Is Regulated by Adr1p and Pip2p-Oaf1p^*

Hanspeter Rottensteiner , Leila Wabnegger  ¶, Ralf Erdmann , Barbara Hamilton , Helmut Ruis  , Andreas Hartig  and Aner Gurvitz  ||

From the Institut für Physiologische Chemie, Ruhr-Universität Bochum, D-44780 Bochum, Germany, and the Max F. Perutz Laboratories, University Departments at the Vienna Biocenter, Department of Biochemistry and Molecular Cell Biology, University of Vienna, and Ludwig Boltzmann-Forschungsstelle für Biochemie, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria

Saccharomyces cerevisiae genes involved in fatty acid degradation contain in their promoters oleate response elements (OREs) and type 1 upstream activation sequences (UAS1s) that bind Pip2p-Oaf1p and Adr1p, respectively. The promoter of the PIP2 gene was found to contain a potential UAS1 that consists of a tandem array of CYCCRR half-sites in an overlapping arrangement with a previously characterized ORE. Electrophoretic mobility shift analysis demonstrated that Adr1p bound to UAS1_PIP2, and Northern analysis in combination with a lacZ reporter gene confirmed that Adr1p influenced the transcription of PIP2. Immunoprecipitation showed that, in adr1 mutant cells grown on oleic acid, Pip2p was less abundant compared with the corresponding wild-type. In addition, the amount of Pip2p-Oaf1p that bound to a target ORE in vitro was reduced in mutant extracts compared with the wild-type. Transcription of the oleic acid-inducible genes SPS19 and CTA1, which rely on both Pip2p-Oaf1p and Adr1p for their regulation, was reduced in adr1 mutant cells. However, by ectopically restoring levels of Pip2p in adr1 cells grown on oleic acid medium, transcription of both genes increased 2-fold compared with the control. This partial suppression of the adr1 mutant phenotype was additionally manifested by moderate utilization of oleic acid. Hence, both the expression as well as the action of the two transcription factors, Adr1p and Pip2p-Oaf1p, are interconnected, which allows for an elaborate control of fatty acid-inducible genes.

Received for publication, April 18, 2003

^* This work was supported by the Fonds zur Förderung der wissenschaftlichen Forschung (Vienna, Austria) Grants P12061-MOB (to H. R. and B. H.) and P14956-B07 (to A. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Current address: Division of Biochemistry and Molecular Biology, Brain Research Inst., University of Vienna, Spitalgasse 4, A-1090 Vienna, Austria. Tel.: 43-1-4277-629-53; Fax: 43-1-4277-629-60.

We dedicate this work to the memory of our co-author, Professor H. Ruis, who died September 1, 2001.

^|| To whom correspondence should be addressed. Tel. 43-1-4277-52815; Fax: 43-1-4277-9528; E-mail: AG{at}abc.univie.ac.at.

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