Both PPREs contribute to the PPAR-dependent transactivation of GSPA and AOX

Copyright information:Taken from "Identification of a Gene Sharing a Promoter and Peroxisome Proliferator-Response Elements With Acyl-CoA Oxidase Gene"PPAR Research 2006;2006():-.Published online 12 Dec 2006PMCID:PMC1779578. (a) Schematic view of the reporter constructs. Upstream and intron regions are depicted with horizontally long boxes, whereas exons with vertically long gray boxes. Nucleotide numbers correspond to those in . The −161/483 region was included in common in both pGSPAluc and pMmAOXluc in opposite orientations. Only parts of 5′ noncoding stretches of exon 2 of GSPA and exon 1 of AOX were included in the respective reporters. In pGSPAluc, whole exon 1 that is noncoding and intron 1 were included, while a 5.1 kb region in the middle of intron 1 was omitted. pMmAOXBluc was also prepared as to lack both PPRE-1 and PPRE-2, but retained the GC-rich region. (b) and (c) Reporter assays for GSPA and AOX expressions, respectively. pGSPAluc, pMmAOXluc, and their mutants involving PPRE-1 (mutP1), PPRE-2 (mutP2), or both (mutP1/mutP2) were transfected into HeLa cells with or without a PPAR expression plasmid, and after transfection, the cells were cultured in the presence or absence of Wy14, 643. For AOX, a minimal promoter vector, pMmAOXBluc, was also employed. Letters “pMm” are omitted from the names of plasmids in the figure. In both (b) and (c), the luciferase activities are shown as relative values, taking the values of respective wild-type constructs in the absence of PPAR expression plasmid and Wy14643, as 1. Mean values of three independent assays are given, together with standard deviations. The actual mean luciferase activity values in the presence of both PPAR and Wy14, 643 were 2.15×10 and 2.37×10 luciferase units for pGSPAluc and pMmAOXluc, respectively.