Published online before print May 20, 2002, doi: 10.1161/01.CIR.0000018127.10968.34
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(Circulation. 2002;105:2760.)
© 2002 American Heart Association, Inc.
Basic Science Reports |
Obligatory Role of Cyclic Adenosine Monophosphate Response Element in Cyclooxygenase-2 Promoter Induction and Feedback Regulation by Inflammatory Mediators
From the Vascular Biology Research Center, Institute of Molecular Medicine and Division of Hematology (Y.Z., M.S., W.-G.D., X.-M.X., K.K.W.), University of Texas Health Science Center, Houston, Tex; and Institut für Pharmakologie und Klinische Pharmakologie (K.S., J.M.-K.), Heinrich-Heine-Universtät Düsseldorf, Düsseldorf, Germany.
Correspondence to Kenneth K. Wu, MD, PhD, Division of Hematology, University of Texas-Houston Medical School, 6431 Fannin, MSB 5.016, Houston, TX 77030. E-mail Kenneth.K.Wu{at}uth.tmc.edu
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-
. 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.
Key Words: prostaglandins • interleukins • endothelium
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