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J. Biol. Chem., Vol. 280, Issue 12, 11192-11202, March 25, 2005

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Chromatin Modification and the Endothelial-specific Activation of the E-selectin Gene^*

Leonard C. Edelstein, Amy Pan, and Tucker Collins

From the Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts 02115

E-selectin plays a role in the binding and extravasation of leukocytes from the bloodstream. The E-selectin gene is rapidly and transiently expressed by endothelial cells activated by inflammatory stimuli. Despite the identification of factors critical for cytokine-induced activation of the E-selectin promoter, little is known about the mechanisms that restrict the gene expression to endothelial cells. We used in vivo approaches to characterize the E-selectin promoter in primary cultures of human umbilical vein endothelial cells and umbilical artery smooth muscle cells. In endothelial cells specifically, nucleosomes are remodeled after tumor necrosis factor (TNF) induction. Chromatin immunoprecipitation analysis demonstrated the binding of the p65 (RelA) component of nuclear factor-B (NF-B) to the endogenous E-selectin promoter after TNF stimulation along with IB kinase . Multiple coactivators, including p300, steroid receptor coactivator-1, and p300/cAMP-response element-binding protein (CREB)-binding protein (CBP)-associated factor localize differentially to the E-selectin promoter. Additionally, TNF induced localized histone hyperacetylation, phosphorylation, and methylation in the E-selectin gene specifically in endothelial cells. Post-induction repression of E-selectin expression is associated with recruitment of multiple deacetylases. Collectively, these studies suggest a model for the selective induction of the E-selectin gene in which the core promoter chromatin architecture is specifically modified in endothelial cells.

Received for publication, November 17, 2004 , and in revised form, January 19, 2005.

^* This work was supported by National Institutes of Health Grant HL-45462. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Pathology, Children's Hospital Boston, 300 Longwood Ave., Boston, MA 02115. Tel.: 617-355-5806; Fax: 617-730-0168, E-mail: tcollins{at}rics.bwh.harvard.edu.

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