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Papers In Press, published online ahead of print September 30, 2002
Department of Medicine, St George Clinical School, Sydney, NSW 2052
Corresponding Author: b.h.chong{at}unsw.edu.au
The exposure of collagen fibres at sites of vascular injury results in the adherence of platelets and their subsequent activation. The platelet collagen receptor glycoprotein (GP) VI plays a crucial role in platelet activation and thrombus formation and decreased levels or defective GPVI may lead to excessive bleeding. In addition, elevated levels of collagen receptors may predispose individuals to coronary heart disease or strokes. GPVI expression is restricted to platelets and their precursor cell, the megakaryocyte (MK). In this study we investigate the regulation of GPVI expression and show that thrombopoietin (TPO) induces its expression in the megakaryocytic cell line UT-7/TPO. A 5'-region flanking the transcription start point of the GPVI gene was cloned (-694 to +29) and we report that this putative GPVI promoter bestows MK specific expression. Deletion analyses and site directed mutagenesis identified Sp1227, GATA177 and Ets48 sites as essential for GPVI expression. We show that transcription factors GATA-1, Fli-1 and Sp1 can bind to and activate this promoter. Finally, GPVI mRNA was detected only in megakaryocytic cell lines expressing both Fli-1 and GATA-1, and we show that over-expression of Fli-1 in a stable cell line (which expresses endogenous GATA-1 and Sp1) results in expression of the endogenous GPVI gene.
J. Biol. Chem, 10.1074/jbc.M206127200
Submitted on June 20, 2002
Revised on September 6, 2002
Accepted on September 30, 2002
Cloning and analysis of the thrombopoietin-induced megakaryocyte-specific glycoprotein VI promoter and its regulation by GATA-1, Fli-1 and Sp1
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