Studies on transcriptional regulation of Cyp3a16 gene in mouse livers by application of direct DNA injection method

Biochem Biophys Res Commun. 2001 Oct 5;287(4):820-4. doi: 10.1006/bbrc.2001.5663.

Abstract

CYP3A16 is expressed in the mouse liver specifically during a fetal and puberty life. Functional regions responsible for the transcriptional regulation of the Cyp3a16 gene were identified by an in vivo direct DNA injection into mouse livers followed by a luciferase assay. The results of the deletion analysis of a 5'-flanking sequence suggested the existence of an adult-specific repressor(s) interacting with the Cyp3a16 gene. In addition, a positive regulatory element was assumed to be present in a region from -146 to -56 of the Cyp3a16 gene. A hepatocyte nuclear factor 4 (HNF-4)-binding consensus sequence was found in this region. In fact, HNF-4alpha bound to this sequence as evidenced by a gel mobility shift assay. The role of the HNF-4-binding sequence was further examined by an introduction of mutations in this sequence. The introduction of the mutations resulted in a reduced activity of a luciferase in the assay. These results obtained by an application of the direct DNA injection method suggest that the HNF-4alpha activates the transcription of the Cyp3a16 gene in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging
  • Animals
  • Aryl Hydrocarbon Hydroxylases*
  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
  • Binding Sites
  • Cytochrome P-450 CYP3A
  • Cytochrome P-450 Enzyme System / genetics*
  • DNA / genetics*
  • DNA / metabolism
  • DNA-Binding Proteins / metabolism
  • Gene Expression Regulation, Enzymologic*
  • Genes, Reporter
  • Hepatocyte Nuclear Factor 4
  • Liver / enzymology
  • Liver / metabolism*
  • Luciferases / genetics
  • Luciferases / metabolism
  • Male
  • Mice
  • Mice, Inbred ICR
  • Microinjections
  • Oxidoreductases, N-Demethylating / genetics*
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Protein Binding
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcription, Genetic / genetics*

Substances

  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
  • DNA-Binding Proteins
  • Hepatocyte Nuclear Factor 4
  • Phosphoproteins
  • Recombinant Fusion Proteins
  • Tcfl4 protein, mouse
  • Transcription Factors
  • DNA
  • Cytochrome P-450 Enzyme System
  • Luciferases
  • Aryl Hydrocarbon Hydroxylases
  • Cytochrome P-450 CYP3A
  • Oxidoreductases, N-Demethylating