Structural and Functional Characterization of the 5′-Flanking Region of the Rat and Human Cytochrome P450 2E1 Genes: Identification of a Polymorphic Repeat in the Human Gene

Abstract

Cytochrome P450 2E1 (CYP2E1) is a toxicologically very important enzyme with a high extent of interindividual variability in expression. We sequenced and characterized the 5′-flanking region of the human and rat CYP2E1 genes. The identity between the human and rat sequences (−3.8 kb to +1 kb) was generally between 35 and 60%, and the most similar regions were found in the proximal part of the sequence. Two more distant regions at −1.6 to −2.0 kb and −2.5 to −2.8 kb in the human sequence were also found to have high identity to the rat sequence. A polymorphic repeat sequence in the human gene was found between −2178 to −1945 bp. The common allele (CYP2E1*1C) contained 6 repeats (each 42–60 bp long) and the rare allele (CYP2E1*1D) had 8 repeats with an allele frequency of 1% among Caucasians and 23% among Chinese. The CYP2E1 5′-flanking regions of the human (−3712 bp to +10 bp) and rat (−3685 bp to +28 bp) genes were ligated in front of a luciferase reporter gene and transfected into rat hepatoma Fao and human hepatoma B16A2 cells. Important species specificity was noted in the control of gene expression and regions of negative and positive cis-acting elements were localized. No difference was seen in the constitutive expression between the two polymorphic forms. The importance of this repeat polymorphism for high and low inducible CYP2E1 phenotypes is discussed.