Interleukin-1 {beta}-induced Id2 gene expression is mediated by Egr-1 in vascular smooth muscle cells

doi:10.1016/j.cardiores.2007.06.015

Cardiovascular Research 2007 76(1):141-148; doi:10.1016/j.cardiores.2007.06.015

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Interleukin-1 β-induced Id2 gene expression is mediated by Egr-1 in vascular smooth muscle cells

Xiaojun Zhu^a^,*^,1, Yiming Lin^b^,d^,1, Methode Bacanamwo^b, Lin Chang^c, Rui Chai^c, Ivana Massud^b, Jifeng Zhang^c, Minerva T. Garcia-Barrio^b, Winston E. Thompson^d and Yuqing E. Chen^c

^aInstitute of Molecular Medicine, Peking University, No. 5 Yi He Yuan Road, Beijing, 100871, P.R. China
^bCardiovascular Research Institute, Morehouse School of Medicine, 720 Westview Dr SW, Atlanta, GA 30310, USA
^cCardiovascular Center, University of Michigan Medical Center, 1150 W. Medical Center Drive, Ann Arbor, MI 48109, USA
^dDepartment of Obstetrics and Gynecology, Cooperative Reproductive Science Research Center, Morehouse School of Medicine, Atlanta, GA 30310, USA

*Corresponding author. Institute of Molecular Medicine, Peking University, No. 5 Yi He Yuan Road, Beijing, PR China 100871. Tel./fax: +86 10 6275 4557. zhuxiaojun{at}pku.edu.cn

Objective Id2 (inhibitor of DNA-binding 2), a member of thehelix–loop–helix family of transcription regulators,plays important roles in cell proliferation and differentiation.Recent reports have documented that Id2 is up-regulated duringvascular lesion formation and overexpression of Id2 promotesvascular smooth muscle cell (VSMC) proliferation. However, thetranscriptional regulation of Id2 gene expression in VSMC remainsunexplored.

Methods and results Using Northern- and Western-blot analyses,we documented that interleukin-1β (IL-1β) inducedId2 gene expression in VSMC in a time- and dose-dependent manner.Overexpression of early growth response-1 (Egr-1) in VSMC inducedId2 expression while IL-1β-induced Id2 expression was abrogatedin VSMC by the Egr-1 repressor, NGFI-A binding protein 2 (NAB2),expressed from an adenovirus. Overexpression of Egr-1 transactivatedthe Id2 promoter in reporter assays dependent on the presenceof intact putative Egr-1 binding sites as determined by mutagenesis.Finally, electrophoretic mobility shift assays (EMSA) demonstratedthat the Egr-1 protein can bind the Egr-1 sites derived fromthe human Id2 promoter in vitro and chromatin immunoprecipitationidentified the putative Egr-1 site between –723 to –712as the functional Egr-1 binding site in vivo.

Conclusions Our data demonstrate that IL-1β-induced Id2expression in VSMC is mediated by the transcription factor Egr-1in VSMC.

KEYWORDS Interleukin-1β; Id2; Egr-1; Vascular smooth muscle cell

¹ X.Z. and Y.L. equally contributed to this paper.

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