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A more recent version of this article appeared on July 18, 2003
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M304097200v1
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Papers In Press, published online ahead of print May 14, 2003
J. Biol. Chem, 10.1074/jbc.M304097200
Submitted on April 18, 2003
Revised on May 14, 2003
Accepted on May 14, 2003

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

Ordinariat I, Vienna University, Vienna A-1030

Corresponding Author: AG{at}abc.univie.ac.at

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 halfsites in an overlapping arrangement with a previously characterized ORE. Electrophoretic mobility shift analysis demonstrated that Adr1p bound to UAS1PIP2, and Northern analysis in combination with a lacZ reporter gene confirmed that Adr1p influenced the transcription of PIP2. Immunoprecipitation showed that in adr1delta 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 adr1delta mutant cells. However, by ectopically restoring levels of Pip2p in adr1delta cells grown on oleic acid medium, transcription of both genes increased twofold compared to the control. This partial suppression of the adr1delta 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.


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