A +220 GATA motif mediates basal but not endotoxin-repressible expression of the von Willebrand factor promoter in Hprt-targeted transgenic mice

J Thromb Haemost. 2009 Aug;7(8):1384-92. doi: 10.1111/j.1538-7836.2009.03501.x. Epub 2009 May 30.


Background: The von Willebrand factor (VWF) gene is a marker for spatial and temporal heterogeneity of the endothelium. A GATA motif at +220 has been implicated in basal VWF expression in vitro. Other studies have shown that GATA3 and VWF are transcriptionally downregulated in response to inflammatory mediators.

Objectives: Our goal was to determine the importance of the +220 GATA motif in mediating expression of VWF promoter in vivo, and to elucidate whether the GATA element plays a role in spatial and/or temporal regulation of VWF expression.

Methods: ChIP and electrophoretic mobility shift assays were carried out in human umbilical vein endothelial cells (HUVEC). Reporter gene constructs containing 3.6 kb of the human VWF promoter with and without a mutation of the +220 GATA element were transfected into cultured endothelial cells or targeted to the Hprt locus of mice. The Hprt-targeted mice were subjected to endotoxemia.

Results: In protein-DNA binding assays, the +220 GATA motif bound GATA-2, -3 and -6. Mutation of the GATA site resulted in reduced basal promoter activity in HUVEC. When targeted to the Hprt locus of mice, the GATA mutation resulted in a significant, proportionate reduction of promoter activity in LacZ expressing vascular beds. Systemic administration of lipopolysaccharide (LPS) resulted in a widespread reduction in VWF mRNA expression and promoter activity. LPS-mediated repression of the VWF promoter was unaffected by the GATA mutation.

Conclusions: A region of the VWF promoter between -2182 and the end of the first intron contains information for LPS-mediated gene repression. The +220 GATA motif is important for basal, but not LPS-repressible expression of the VWF gene.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cells, Cultured
  • Down-Regulation / genetics
  • Endothelium, Vascular / cytology
  • GATA Transcription Factors / physiology*
  • Gene Expression Regulation / drug effects*
  • Humans
  • Hypoxanthine Phosphoribosyltransferase / genetics
  • Lipopolysaccharides / pharmacology*
  • Mice
  • Mice, Transgenic
  • Promoter Regions, Genetic
  • RNA, Messenger / analysis
  • von Willebrand Factor / genetics*


  • GATA Transcription Factors
  • Lipopolysaccharides
  • RNA, Messenger
  • von Willebrand Factor
  • Hypoxanthine Phosphoribosyltransferase