Regulation of the yeast DPP1-encoded diacylglycerol pyrophosphate phosphatase by transcription factor Gis1p.
The Saccharomyces cerevisiae DPP1-encoded diacylglycerol pyrophosphate phosphatase catalyzes the dephosphorylation of diacylglycerol pyrophosphate to form phosphatidate and Pi. The enzyme also dephosphorylates phosphatidate to form diacylglycerol and Pi. Because diacylglycerol pyrophosphate, phosphatidate, and diacylglycerol have roles as lipid signal molecules in higher eukaryotic cells, it is important to understand how diacylglycerol pyrophosphate phosphatase is regulated. Analysis of DPP1 expression using PDPP1-lacZ reporter genes with a series of deletions from the 5' end of the promoter indicated sequences responsible for enzyme expression. Three binding sites (URSPDS) for transcription factor Gis1p were identified in the DPP1 promoter (consensus sequence of 5'-T(A/T)AGGGAT-3'). A gis1 Delta mutant exhibited elevated levels of DPP1 expression and diacylglycerol pyrophosphate phosphatase activity. Direct interaction between Gis1p and DPP1 promoter elements was demonstrated by electrophoretic mobility shift assays. Mutations in the three URSPDS elements within the DPP1 promoter abolished Gis1p-DNA interactions in vitro and abolished the regulation of DPP1 in vivo. These data indicated that Gis1p was a repressor of DPP1 expression. Phospholipid composition analysis of the gis1 Delta mutant showed that Gis1p played a role in regulating the cellular level of diacylglycerol pyrophosphate, as well as the levels of the major phospholipids phosphatidylethanolamine and phosphatidylcholine.
Oshiro
June
J
Graduate Program in Microbiology and Molecular Genetics and Deparetment of Food Science, Rutgers University, New Brunswick, New Jersey 08901, USA.
Han
Gil-Soo
GS
Iwanyshyn
Wendy M
WM
Conover
Kristi
K
Carman
George M
GM
D000595:Amino Acid Sequence; D001483:Base Sequence; D024202:Electrophoretic Mobility Shift Assay; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005800:Genes, Fungal; D017930:Genes, Reporter; D056466:Histone Demethylases; D008969:Molecular Sequence Data; D009154:Mutation; D011755:Pyrophosphatases / Q000235:genetics*; D012097:Repressor Proteins / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics
D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C400297:GIS1 protein, S cerevisiae; D056466:Histone Demethylases; D001616:beta-Galactosidase; D011755:Pyrophosphatases; C097892:diacylglycerol pyrophosphate phosphatase