Molecular Mechanism of Basal CYP3A4 Regulation by Hepatocyte Nuclear Factor 4α: Evidence for Direct Regulation in the Intestine
| Autor: | Heike Tegude, Anke Schnabel, Kathrin Klein, Oliver Burk, Ulrich M. Zanger, Michel Eichelbaum |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Chicken ovalbumin upstream promoter-transcription factor
Pharmaceutical Science Biology Response Elements digestive system Cytochrome P-450 Enzyme System Cell Line Tumor medicine Cytochrome P-450 CYP3A Humans RNA Messenger Intestinal Mucosa Binding site Promoter Regions Genetic Receptor Transcription factor Pharmacology Binding Sites Molecular biology Hepatocyte nuclear factors Hepatocyte Nuclear Factor 4 Liver Hepatocyte nuclear factor 4 Nuclear receptor Mechanism of action medicine.symptom |
| Zdroj: | Drug Metabolism and Disposition. 35:946-954 |
| ISSN: | 1521-009X 0090-9556 |
| DOI: | 10.1124/dmd.106.013565 |
| Popis: | Cytochrome P450 3A4 plays an outstanding role in the metabolism of clinically used drugs and shows a marked interindividual variability in expression even in the absence of inducing agents. Thus, regulation of basal expression contributes considerably to variability. The nuclear receptor hepatocyte nuclear factor 4alpha (HNF4alpha) was previously shown to be associated with basal hepatic CYP3A4 expression. As how HNF4alpha regulates basal expression of CYP3A4 still remains elusive, we systematically screened 12.5 kilobase pairs (kb) of the CYP3A4 5' upstream region for activation by the receptor in the human intestinal cell line LS174T. In this study, we newly identified two widely separated regions mediating the activation by HNF4alpha: a far distal region at -9.0 kb and the proximal promoter region at approximately -0.2 kb. By gel shift experiments and transient transfections, we characterized direct repeat (DR) 1-type motifs in both regions as functional HNF4alpha response elements. Cooperation of the two regions was shown to be required for maximal activation by HNF4alpha. The effect of HNF4alpha was antagonized by chicken ovalbumin upstream promoter transcription factor II, which was shown to bind to one of the DR1 motifs. Furthermore, activation of CYP3A4 via the DR1 element in the proximal promoter depends on an additional, yet unknown, factor, which is binding at approximately -189 base pairs. Physiological relevance of this position for activation by HNF4alpha in vivo is suggested by the presence of a binding activity in small intestine similar to that in LS174T cells. In summary, we here have elucidated a molecular mechanism of direct regulation of CYP3A4 by HNF4alpha, which is probably specific for the intestine. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|