Transcriptional regulation of the human soluble epoxide hydrolase gene EPHX2.
Soluble epoxide hydrolase (sEH) is a multifunctional protein encoded by the EPHX2 gene. The biological functions and enzyme kinetics of sEH have been extensively investigated, however, little is known about its transcriptional regulation and mechanisms of tissue specific expression. Here, a luciferase gene based reporter assay was used to identify the minimal promoter and cis regulatory elements of EPHX2. The minimal promoter was identified as a GC-rich region between nts -374 and +28 with respect to the putative transcriptional start site. A reporter plasmid carrying this minimal promoter showed higher or similar activities in comparison to a reporter plasmid carrying nts -5,974 to +28 of EPHX2 in 9 human cell lines that were tested. Sp1 binding sites that are involved in augmenting the minimal promoter activity of EPHX2 were identified by nested deletion analysis, site-specific mutation, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay.
Tanaka
Hiromasa
H
Department of Entomology and the Cancer Research Center, University of California, Davis, CA 95616, USA.
Kamita
Shizuo G
SG
Wolf
Nicola M
NM
Harris
Todd R
TR
Wu
Zhaoju
Z
Morisseau
Christophe
C
Hammock
Bruce D
BD
D024506:5' Flanking Region; D001483:Base Sequence; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D003582:Cytogenetics; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D004851:Epoxide Hydrolases / Q000235:genetics*; D020862:GC Rich Sequence; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
D017931:DNA Primers; D004851:Epoxide Hydrolases; C506428:EPHX2 protein, human