Suppression of CYP2C11 gene transcription by interleukin-1 mediated by NF-kappaB binding at the transcription start site

Arch Biochem Biophys. 2000 May 1;377(1):187-94. doi: 10.1006/abbi.2000.1772.


Inflammatory cytokines cause the down-regulation of multiple cytochrome P450 mRNAs, but the transcriptional mechanisms involved are not known. We investigated the role of a putative negative NF-kappaB-responsive element, nkappaB-RE1, in the down-regulation of the CYP2C11 gene in rat hepatocytes. This sequence spans the transcription start site of CYP2C11, from positions -2 to +8. Electrophoretic mobility shift assays showed that nuclear extracts from livers of rats treated with bacterial lipopolysaccharide, or from hepatocytes treated with interleukin-1beta, formed a protein complex with an oligonucleotide probe containing the nkappaB-RE1, and that this complex contained predominantly the p50 subunit of NF-kappaB. Binding of NF-kappaB to the nkappaB-RE1 probe was of lower affinity than to a probe containing the prototypic NF-kappaB enhancer of the immunoglobulin kappa chain gene. Mutations in the 5'-end of the nkappaB-RE1, and to a lesser extent the 3'-end, reduced the affinity of NF-kappaB for this element. Introduction of the 5'-mutation into nkappaB-RE1 abolished the response of the -200-CYP2C11-chloramphenicol acetyltransferase reporter construct to interleukin-1 or lipopolysaccharide. We conclude that nkappaB-RE1 is a functional negative regulatory element that participates in the inflammatory suppression of CYP2C11.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Aryl Hydrocarbon Hydroxylases*
  • Base Sequence
  • Binding, Competitive
  • Cell Nucleus / chemistry
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Consensus Sequence / genetics
  • Cytochrome P-450 Enzyme System / genetics*
  • Cytochrome P450 Family 2
  • DNA / genetics
  • DNA / metabolism*
  • Down-Regulation / drug effects*
  • Interleukin-1 / pharmacology*
  • Lipopolysaccharides / pharmacology
  • Liver / cytology
  • Liver / drug effects
  • Liver / metabolism
  • Male
  • Molecular Weight
  • Mutation / genetics
  • NF-kappa B / chemistry
  • NF-kappa B / metabolism*
  • Promoter Regions, Genetic / genetics
  • Protein Binding
  • Rats
  • Rats, Sprague-Dawley
  • Response Elements / genetics
  • Steroid 16-alpha-Hydroxylase*
  • Steroid Hydroxylases / genetics*
  • Transcription, Genetic / drug effects*
  • Transcription, Genetic / genetics


  • Interleukin-1
  • Lipopolysaccharides
  • NF-kappa B
  • DNA
  • Cytochrome P-450 Enzyme System
  • Steroid Hydroxylases
  • Aryl Hydrocarbon Hydroxylases
  • CYP2C11 protein, rat
  • Cytochrome P450 Family 2
  • Steroid 16-alpha-Hydroxylase