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J. Biol. Chem., Vol. 283, Issue 16, 10264-10275, April 18, 2008

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Binding Characteristics and Regulatory Mechanisms of the Transcription Factors Controlling Oleate-responsive Genes in Saccharomyces cerevisiae^*

From the Department of Biology, City College of the City University of New York, New York, New York 10031

Transcriptional activation of many genes involved in peroxisome-related functions is regulated by the Oaf1p, Pip2p, and Adr1p transcription factors in Saccharomyces cerevisiae. We have analyzed the in vivo binding characteristics of Oaf1p-Pip2p and found that this complex is recruited to its target oleate-response element (ORE) under all growth conditions tested. In addition, this complex also binds to ORE-containing genes that do not appear to be regulated by these proteins, as well as to some genes lacking conventional OREs. The recruitment of the Oaf1p-Pip2p complex was greatly increased upon glucose derepression, possibly due to Oaf1p phosphorylation with only moderate increases upon oleate induction. Thus, this complex may receive a nutritional cue while it is already bound to DNA, suggesting that, in addition to the increase in Oaf1p-Pip2p binding, other mechanism(s) such as enhanced Adr1p association may drive the expression of highly inducible fatty acid-responsive genes. Adr1p binds to target genes in an oleate-dependent fashion and is involved in Oaf1p-Pip2p binding. In turn, the Oaf1p-Pip2p complex appears to be important for Adr1p binding to a subset of oleate-responsive genes. Adr1p is a positive regulator of ORE-containing genes, but it also acts as a negative factor in expression of some of these genes. Finally, we have also shown that Adr1p is directly involved in mediating oleate induction of Oaf1p-Pip2p target genes.

Received for publication, October 3, 2007 , and in revised form, February 5, 2008.

^* This work was supported by American Heart Association Grant 0350364N, NIGMS Grant SO6 GM-08168 from the National Institutes of Health, and National Institutes of Health Grant RCMI RR 03060. 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.

¹ Present address: Centre of Bioengineering, Russian Academy of Sciences, Moscow 117984, Russia.

² Present address: Cardiovascular, Metabolic, and Endocrine Diseases Department, Pfizer Inc., Groton, CT 06385.

³ To whom correspondence should be addressed: 535 East 80th St., New York, NY 10075. Tel.: 212-794-5417; Fax: 212-794-5378; E-mail: Gillian.small{at}mail.cuny.edu.

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