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Originally published In Press as doi:10.1074/jbc.M108075200 on December 10, 2001

J. Biol. Chem., Vol. 277, Issue 9, 6923-6928, March 1, 2002
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Dynamic Regulation of Cyclooxygenase-2 Promoter Activity by Isoforms of CCAAT/Enhancer-binding Proteins*

Ying Zhu, Michael A. Saunders, Howard Yeh, Wu-guo Deng, and Kenneth K. WuDagger

From the Vascular Biology Research Center, Institute of Molecular Medicine, and Division of Hematology, University of Texas-Houston Medical School, Houston, Texas 77030

To elucidate the mechanism by which isoforms of CCAAT/enhancer-binding proteins regulate cyclooxygenase-2 expression, we determined by a novel technique binding of six isoforms of this transactivator to two sequence-specific CCAAT/enhancer-binding protein (-132/-125) and cyclic AMP (-59/-53) regulatory elements in human foreskin fibroblasts treated with phorbol 12-myristate 13-acetate for 4 h. The delta  isoform bound to these two elements at basal state, which was displaced by full-length as well as two truncated beta  isoforms, a 41-kDa liver-enriched activating protein and a 16-kDa liver-enriched inhibitory protein, after phorbol ester stimulation. Kinetic analysis shows time-dependent changes in beta  and delta  binding that were concordant with time-dependent increase in cyclooxygenase-2 induction. Overexpression of the 16-kDa beta  isoform blocked the promoter activity and protein level induced by phorbol ester. Paradoxically, it increased binding of beta  isoforms to the sequence-specific promoter DNA but suppressed cyclooxygenase-2 promoter activation by p300 cotransfection. These findings provide new insight into the regulation of cyclooxygenase-2 promoter by an interplay between two opposite beta  isoforms and p300 co-activator.

* This work was supported by Grant R01 HL-50675 from the National Heart, Lung and Blood Institute and the NINDS, National Institutes of Health Grant P50 NS-23327.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Vascular Biology Research Center and Div. of Hematology, Univ. of Texas-Houston Medical School, 6431 Fannin, MSB 5.016, Houston, TX 77030. Tel.: 713-500-6801; Fax: 713-500-6812; E-mail: Kenneth.K.Wu@uth.tmc.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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