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Molecular mechanisms involved in the regulation of the cytochrome P450 epoxygenase CYP2J5
Author(s) -
Graves Joan P,
Zhang Donghui,
Maestro Miguel A,
Servitja Joan M,
King Lorraine M,
Swope Deborah L,
Korach Kenneth S,
Ferrer Jorge,
Zeldin Darryl C
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.479.2
Subject(s) - transfection , microbiology and biotechnology , biology , chromatin immunoprecipitation , androgen receptor , reporter gene , promoter , gene , binding site , gene expression , genetics , prostate cancer , cancer
CYP2J5 is expressed in kidney and liver, and metabolizes arachidonic acid to epoxyeicosatrienoic acids. Herein, we identify the molecular mechanisms responsible for regulation of this gene. The transcriptional start site of the Cyp2j5 gene was located and analysis of the proximal promoter region identified a putative HNF1alpha binding site and a putative androgen response element (ARE). Electrophoretic mobility shift assays (EMSAs) demonstrated binding of androgen receptor (AR) to the Cyp2j5 ARE. Chromatin immunoprecipitation assays with mouse kidney cells confirmed specific binding of AR to the Cyp2j5 ARE, and this was increased in cells treated with androgen compared to cells treated with vehicle. EMSAs also showed the binding of HNF1alpha to the putative Cyp2j5 binding site. Reporter assays with a Cyp2j5 promoter vector demonstrated activation when an HNF1alpha overexpression vector was co‐transfected but not when there was a mutation in the HNF1alpha site. Immunohistochemistry demonstrated a marked reduction in the expression of Cyp2j5 in livers but only minimal changes in kidneys of HNF1alpha knockout mice relative to wild type controls. These findings provide a molecular basis for the observed gender and tissue specific differences in the expression of the Cyp2j5 gene. This research was supported by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences.