Structure and Regulation of the Ethanol-Inducible Cytochrome P450j

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Abstract
Specific polyclonal antisera against microsomal ethanol-inducible cytochrome P450 (P450j, P450IIE) were prepared and utilized to isolate cDNA for P450j from lambda gt11 cDNA libraries. The longest cDNAs encoding P450j of rat and human were completely sequenced. The rat P450j sequence was compared to those of other P450s (P450II gene family members) to determine the structural similarity. Southern-blot analysis of rat and human genomic DNAs verified that only a single gene shared extensive homology with P450j. Cloned P450j cDNA and antibodies were used to study the expression of P450j gene during development and by various inducers as well as in pathological conditions. By combination of cDNA hybridization and immunoblot analyses, three types of P450j gene expression were observed: transcriptional activation during development; post-transcriptional activation (probably via protein stabilization) by various inducers such as pyrazole, 4-methylpyrazole, acetone, and ethanol; and mRNA stabilization in diabetic and starved animals. These three different types of P450j induction appeared to be present not only in liver but also in lung and kidney tissues.

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