Influence of a GT repeat element on shear stress responsiveness of the VWF gene promoter.

Abstract

Plasma von Willebrand factor (VWF) is mainly derived from endothelial cells, cells that express a large repertoire of genes that are transcriptionally regulated by fluid shear stress. Endothelial VWF expression is not uniform throughout the vasculature, and levels are increased at regions associated with disturbed blood flow and steep gradients of shear stress. It is, however, unknown whether shear stress influences the regulation of VWF gene expression. Our objective was to evaluate the effect of shear stress on endogenous endothelial VWF mRNA expression and VWF promoter (-2722 to -1224) activity and to determine whether genetic elements modulate this flow-induced expression. A parallel plate flow chamber was used to expose endothelial cells to a shear level of 15 dynes cm(-2) for 24 or 6 h. VWF mRNA expression was analyzed. Various VWF promoter constructs that each contain either SNP haplotypes 1 or 2 and either a 17-GT or a 23-GT repeat element were transfected into endothelial cells, and flow-induced promoter activation was assessed. When endothelial cells were exposed to shear stress, endogenous VWF mRNA expression increased 1.84-fold and average VWF promoter activity was enhanced 3.4-fold. Single nucleotide polymorphisms at -2708 and -2525, and the shear stress-response element at -1585, are not responsible for the shear stress-induced increase. Rather a GT repeat element at -2124 mediates the increase in activity, and the length of this polymorphic repeat element influences the magnitude of induction. Shear stress enhances VWF promoter activity and a polymorphic GT repeat element mediates the stress-induced transactivation.