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Eukaryotic Cell, March 2005, p. 604-614, Vol. 4, No. 3
1535-9778/05/$08.00+0     doi:10.1128/EC.4.3.604-614.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Genomic Analysis of PIS1 Gene Expression

Mary E. Gardocki, Margaret Bakewell, Deepa Kamath, Kelly Robinson, Kathy Borovicka, and John M. Lopes*

Department of Biological Sciences, Wayne State University, Detroit, Michigan

Received 18 September 2004/ Accepted 20 December 2004

The Saccharomyces cerevisiae PIS1 gene is essential and required for the final step in the de novo synthesis of phosphatidylinositol. Transcription of the PIS1 gene is uncoupled from the factors that regulate other yeast phospholipid biosynthetic genes. Most of the phospholipid biosynthetic genes are regulated in response to inositol and choline via a regulatory circuit that includes the Ino2p:Ino4p activator complex and the Opi1p repressor. PIS1 is regulated in response to carbon source and anaerobic growth conditions. Both of these regulatory responses are modest, which is not entirely surprising since PIS1 is essential. However, even modest regulation of PIS1 expression has been shown to affect phosphatidylinositol metabolism and to affect cell cycle progression. This prompted the present study, which employed a genomic screen, database mining, and more traditional promoter analysis to identify genes that affect PIS1 expression. A screen of the viable yeast deletion set identified 120 genes that affect expression of a PIS1-lacZ reporter. The gene set included several peroxisomal genes, silencing genes, and transcription factors. Factors suggested by database mining, such as Pho2 and Yfl044c, were also found to affect PIS1-lacZ expression. A PIS1 promoter deletion study identified an upstream regulatory sequence element that was required for carbon source regulation located downstream of three previously defined upstream activation sequence elements. Collectively, these studies demonstrate how a collection of genomic and traditional strategies can be implemented to identify a set of genes that affect the regulation of an essential gene.

* Corresponding author. Mailing address: Department of Biological Sciences, Wayne State University, 5047 Gullen Mall, Detroit, MI 48202. Phone: (313) 993-7816. Fax: (313) 577-6891. E-mail: jlopes{at}sun.science.wayne.edu.

Eukaryotic Cell, March 2005, p. 604-614, Vol. 4, No. 3
1535-9778/05/$08.00+0     doi:10.1128/EC.4.3.604-614.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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