Expression of the Yeast PIS1 Gene Requires Multiple Regulatory Elements Including a Rox1p Binding Site

• Published in: The Journal of biological chemistry 2003-10, Vol.278 (40), p.38646-38652
• Main Authors: Gardocki, Mary Elizabeth, Lopes, John M.
• Format: Article
• Language: English
• Subjects: Base Sequence; Binding Sites; Carbon - chemistry; Chloramphenicol O-Acetyltransferase - metabolism; Choline - chemistry; Chromatography, Thin Layer; Conserved Sequence; DNA - metabolism; DNA, Complementary - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - metabolism; Gene Deletion; Gene Expression Regulation, Fungal; Genes, Reporter; Hypoxia; Inositol - chemistry; Lipid Metabolism; Models, Biological; Models, Genetic; Molecular Sequence Data; Oxygen - metabolism; Phospholipids - chemistry; Plasmids - metabolism; Promoter Regions, Genetic; Protein Binding; Repressor Proteins - chemistry; Repressor Proteins - metabolism; RNA - metabolism; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins; Transcription, Genetic; Transferases (Other Substituted Phosphate Groups) - chemistry; Transferases (Other Substituted Phosphate Groups) - genetics
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M305251200

The PIS1 gene is required for de novo synthesis of phosphatidylinositol (PI), an essential phospholipid in Saccharomyces cerevisiae. PIS1 gene expression is unusual because it is uncoupled from the other phospholipid biosynthetic genes, which are regulated in response to inositol and choline. Relatively little is known about regulation of transcription of the PIS1 gene. We reported previously that PIS1 transcription is sensitive to carbon source. To further our understanding of the regulation of PIS1 transcription, we carried out a promoter deletion analysis that identified three regions required for PIS1 gene expression (upstream activating sequence (UAS) elements 1-3). Deletion of either UAS1 or UAS2 resulted in an ∼45% reduction in expression, whereas removal of UAS3 yielded an 84% decrease in expression. A comparison of promoters among several Saccharomyces species shows that these sequences are highly conserved. Curiously, the UAS3 element region (-149 to -138) includes a Rox1p binding site. Rox1p is a repressor of hypoxic genes under aerobic growth conditions. Consistent with this, we have found that expression of a PIS1-cat reporter was repressed under aerobic conditions, and this repression was dependent on both Rox1p and its binding site. Furthermore, PI levels were elevated under anaerobic conditions. This is the first evidence that PI levels are affected by regulation of PIS1 transcription.