The transcription factor Erg controls endothelial cell quiescence by repressing activity of nuclear factor (NF)-κB p65

J Biol Chem. 2012 Apr 6;287(15):12331-42. doi: 10.1074/jbc.M112.346791. Epub 2012 Feb 15.

Abstract

The interaction of transcription factors with specific DNA sequences is critical for activation of gene expression programs. In endothelial cells (EC), the transcription factor NF-κB is important in the switch from quiescence to activation, and is tightly controlled to avoid excessive inflammation and organ damage. Here we describe a novel mechanism that controls the activation of NF-κB in EC. The transcription factor Erg, the most highly expressed ETS member in resting EC, controls quiescence by repressing proinflammatory gene expression. Focusing on intercellular adhesion molecule 1(ICAM)-1 as a model, we identify two ETS binding sites (EBS -118 and -181) within the ICAM-1 promoter required for Erg-mediated repression. We show that Erg binds to both EBS -118 and EBS -181, the latter located within the NF-κB binding site. Interestingly, inhibition of Erg expression in quiescent EC results in increased NF-κB-dependent ICAM-1 expression, indicating that Erg represses basal NF-κB activity. Erg prevents NF-κB p65 from binding to the ICAM-1 promoter, suggesting a direct mechanism of interference. Gene set enrichment analysis of transcriptome profiles of Erg and NF-κB-dependent genes, together with chromatin immunoprecipitation (ChIP) studies, reveals that this mechanism is common to other proinflammatory genes, including cIAP-2 and IL-8. These results identify a role for Erg as a gatekeeper controlling vascular inflammation, thus providing an important barrier to protect against inappropriate endothelial activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Binding, Competitive
  • Cells, Cultured
  • DNA / chemistry
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Profiling
  • Gene Expression Regulation*
  • Genes, Reporter
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Human Umbilical Vein Endothelial Cells / physiology*
  • Humans
  • Intercellular Adhesion Molecule-1 / genetics
  • Intercellular Adhesion Molecule-1 / metabolism
  • Luciferases, Renilla / biosynthesis
  • Luciferases, Renilla / genetics
  • Promoter Regions, Genetic
  • Protein Binding
  • Resting Phase, Cell Cycle
  • Trans-Activators / chemistry
  • Trans-Activators / metabolism
  • Trans-Activators / physiology*
  • Transcription Factor RelA / metabolism*
  • Transcription Initiation Site
  • Transcription, Genetic
  • Transcriptional Regulator ERG

Substances

  • ERG protein, human
  • Trans-Activators
  • Transcription Factor RelA
  • Transcriptional Regulator ERG
  • Intercellular Adhesion Molecule-1
  • DNA
  • Luciferases, Renilla