Regulation of the CYP3A4 and CYP3A7 promoters by members of the nuclear factor I transcription factor family.
Despite the established interindividual variability and ontogeny of the CYP3A enzymes, the most abundant phase I drug-metabolizing enzymes in human liver and intestine, the mechanisms that regulate basal expression remain poorly understood. Electrophoretic mobility shift assays using nuclear proteins extracted from human prenatal and postnatal liver samples identified multiple, developmentally distinct nuclear factor I (NFI)-containing protein complexes from human liver bound to sequences from the CYP3A4 (-243/-220) and CYP3A7 (-242/-219) proximal promoters. In addition, a hepatocyte nuclear factor (HNF) 3gamma-containing complex from prenatal liver interacted with CYP3A7-242/-219 but not CYP3A4-243/-220. Cotransfection of HepG2 cells with a CYP3A4 proximal promoter construct and expression vectors for the NFI isoforms NFIA1.1, NFIB2, NFIC1, NFIC2, and NFIX1 enhanced the expression of luciferase activity. In contrast, cotransfection of NFIB2, NFIC1, NFIC2, NFIX1, and NFIX2 reduced the expression of luciferase under the control of the CYP3A7 gene promoter. Mutagenesis of the NFI/HNF3gamma binding sites in the CYP3A7 and CYP3A4 proximal promoters suggests that regulation of basal promoter activity by members of the NFI transcription factor family occur via multiple mechanisms. These results demonstrate that members of the NFI transcription factor family regulate CYP3A4 and CYP3A7 basal expression in an isoform- and promoter-specific manner.
Riffel
Amanda K
AK
Division of Pediatric Clinical Pharmacology and Medical Toxicology, Department of Pediatrics, Children's Mercy Hospital and Clinics, Kansas City, MO 64108, USA.
Schuenemann
Eric
E
Vyhlidal
Carrie A
CA
D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D051544:Cytochrome P-450 CYP3A / Q000235:genetics* / Q000378:metabolism; D005333:Fetus; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008099:Liver / Q000502:physiology; D005810:Multigene Family / Q000502:physiology*; D051839:NFI Transcription Factors / Q000502:physiology*; D011401:Promoter Regions, Genetic
D051839:NFI Transcription Factors; D001189:Aryl Hydrocarbon Hydroxylases; C103123:CYP3A7 protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human