Obligatory role of cyclic adenosine monophosphate response element in cyclooxygenase-2 promoter induction and feedback regulation by inflammatory mediators

Circulation. 2002 Jun 11;105(23):2760-5. doi: 10.1161/01.cir.0000018127.10968.34.

Abstract

Background: Cyclooxygenase-2 (COX-2) plays a key role in human inflammatory disorders such as vascular inflammation. COX-2 promoter activity is induced by proinflammatory mediators, but the role of cyclic adenosine monophosphate response element (CRE) in promoter stimulation remains unclear.

Methods and results: Transient transfection of a 0.9-kb COX-2 promoter fragment bearing CRE mutation abrogated COX-2 promoter activity induced by proinflammatory mediators in human endothelial cells and fibroblasts. Dual mutations of CRE and an upstream CCAAT/enhancer binding protein (C/EBP) site did not have an additional effect. Binding of CREB-2, ATF-2, USF-2, and c-Jun transactivators to a wild-type and CRE-mutated oligonucleotide was analyzed by a novel DNA-binding assay. CREB-2 and ATF-2 in nuclear extracts of unstimulated endothelial cells bound to CRE, whereas USF-2 and c-Jun or c-Fos bound to non-CRE sites. CREB-2 and c-Fos binding was increased by phorbol 12-myristate 13-acetate but not tumor necrosis factor-alpha. The binding assay and chromatin immunoprecipitation revealed binding of P300 coactivator to the COX-2 promoter region.

Conclusions: CRE plays an obligatory role in COX-2 promoter activation by diverse stimuli. CREB-2 and ATF-2 bound to CRE serve as an anchor for P300 interaction with upstream transactivators and downstream transcription machinery.

Publication types

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

MeSH terms

  • Activating Transcription Factor 4
  • Cell Line, Transformed
  • Cells, Cultured
  • Chromatin / metabolism
  • Cyclic AMP / analogs & derivatives
  • Cyclic AMP / metabolism
  • Cyclooxygenase 2
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / enzymology*
  • Endothelium, Vascular / metabolism
  • Enzyme Induction
  • Humans
  • Inflammation Mediators / pharmacology*
  • Interleukin-1 / pharmacology
  • Isoenzymes / genetics*
  • Membrane Proteins
  • Nuclear Proteins / metabolism
  • Promoter Regions, Genetic
  • Prostaglandin-Endoperoxide Synthases / genetics*
  • Prostaglandins / pharmacology
  • Response Elements*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Trans-Activators / metabolism*
  • Transcription Factors / metabolism
  • Transcriptional Activation
  • Tumor Necrosis Factor-alpha / pharmacology
  • Up-Regulation

Substances

  • ATF4 protein, human
  • Chromatin
  • Inflammation Mediators
  • Interleukin-1
  • Isoenzymes
  • Membrane Proteins
  • Nuclear Proteins
  • Prostaglandins
  • Trans-Activators
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • Activating Transcription Factor 4
  • Cyclic AMP
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • Tetradecanoylphorbol Acetate