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Papers In Press, published online ahead of print March 10, 2004
RCAST, University of Tokyo, Tokyo, Tokyo 153-8904
Corresponding Author: minami{at}med.rcast.u-tokyo.ac.jp
Recent evidence supports a role for GATA transcription factors as important signal intermediates in differentiated endothelial cells. The goal of this study was to identify proteins that interact with endothelial-derived GATA transcription factors. Using yeast two-hybrid screening, we identified hematopoietically expressed homeobox (Hex) as a GATA-binding partner in endothelial cells. The physical association between Hex and GATA was confirmed with immunoprecipitation in cultured cells. Hex overexpression resulted in decreased flk-1/KDR expression, both at the level of the promoter and the endogenous gene, and attenuated VEGF-mediated tube formation in primary endothelial cell cultures. In electrophoretic mobility shift assays, Hex inhibited the binding of GATA-2 to the flk-1/KDR 5-UTR GATA motif. Finally, in RNase protection assays, TGF-
J. Biol. Chem, 10.1074/jbc.M308730200
Submitted on August 7, 2003
Revised on March 10, 2004
Accepted on March 10, 2004
Interaction between Hex and GATA transcription factors in vascular endothelial cells inhibits flk-1/KDR-mediated VEGF signaling
1, which has been previously shown to decrease Flk-1 expression by interfering with GATA binding activity, was shown to increase Hex expression in endothelial cells. Taken together, the present study provides evidence for a novel association between Hex and GATA, and suggests that TGF--mediated repression of flk-1/KDR and VEGF signaling involves the inducible formation of inhibitory Hex-GATA complexes.
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