title: Interleukin-1 beta-induced Id2 gene expression is mediated by Egr-1 in vascular smooth muscle cells.
abstract: OBJECTIVE
  Id2 (inhibitor of DNA-binding 2), a member of the helix-loop-helix family of transcription regulators, plays important roles in cell proliferation and differentiation. Recent reports have documented that Id2 is up-regulated during vascular lesion formation and overexpression of Id2 promotes vascular smooth muscle cell (VSMC) proliferation. However, the transcriptional regulation of Id2 gene expression in VSMC remains unexplored.
  
  
  RESULTS
  Using Northern- and Western-blot analyses, we documented that interleukin-1beta (IL-1beta) induced Id2 gene expression in VSMC in a time- and dose-dependent manner. Overexpression of early growth response-1 (Egr-1) in VSMC induced Id2 expression while IL-1beta-induced Id2 expression was abrogated in VSMC by the Egr-1 repressor, NGFI-A binding protein 2 (NAB2), expressed from an adenovirus. Overexpression of Egr-1 transactivated the Id2 promoter in reporter assays dependent on the presence of intact putative Egr-1 binding sites as determined by mutagenesis. Finally, electrophoretic mobility shift assays (EMSA) demonstrated that the Egr-1 protein can bind the Egr-1 sites derived from the human Id2 promoter in vitro and chromatin immunoprecipitation identified the putative Egr-1 site between -723 to -712 as the functional Egr-1 binding site in vivo.
  
  
  CONCLUSIONS
  Our data demonstrate that IL-1beta-induced Id2 expression in VSMC is mediated by the transcription factor Egr-1 in VSMC.
authors.affiliation: Institute of Molecular Medicine, Peking University, No. 5 Yi He Yuan Road, Beijing, 100871, PR China. zhuxiaojun@pku.edu.cn
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhu
authors.forename: Yiming
authors.initials: Y
authors.lastname: Lin
authors.forename: Methode
authors.initials: M
authors.lastname: Bacanamwo
authors.forename: Lin
authors.initials: L
authors.lastname: Chang
authors.forename: Rui
authors.initials: R
authors.lastname: Chai
authors.forename: Ivana
authors.initials: I
authors.lastname: Massud
authors.forename: Jifeng
authors.initials: J
authors.lastname: Zhang
authors.forename: Minerva T
authors.initials: MT
authors.lastname: Garcia-Barrio
authors.forename: Winston E
authors.initials: WE
authors.lastname: Thompson
authors.forename: Yuqing E
authors.initials: YE
authors.lastname: Chen
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000704:Analysis of Variance; D001665:Binding Sites; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D004305:Dose-Response Relationship, Drug; D051766:Early Growth Response Protein 1 / Q000037:antagonists & inhibitors / Q000235:genetics*; D024202:Electrophoretic Mobility Shift Assay; D015870:Gene Expression; D005786:Gene Expression Regulation; D006801:Humans; D051797:Inhibitor of Differentiation Protein 2 / Q000235:genetics*; D053583:Interleukin-1beta / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular; D016297:Mutagenesis, Site-Directed; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D013268:Stimulation, Chemical; D015533:Transcriptional Activation
chemical_list: C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; C497450:ID2 protein, human; D051797:Inhibitor of Differentiation Protein 2; D053583:Interleukin-1beta; C491000:NAB2 protein, human; D012333:RNA, Messenger; D012097:Repressor Proteins