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Originally published In Press as doi:10.1074/jbc.M008798200 on November 29, 2000

J. Biol. Chem., Vol. 276, Issue 7, 5395-5402, February 16, 2001
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Transforming Growth Factor-beta 1-mediated Inhibition of the flk-1/KDR Gene Is Mediated by a 5'-Untranslated Region Palindromic GATA Site*

Takashi MinamiDagger §, Robert D. RosenbergDagger §, and William C. AirdDagger

From the Dagger  Department of Molecular Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215 and the § Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

The angiogenic effects of vascular endothelial growth factor are mediated predominantly by the FLK-1/KDR receptor. An understanding of the transcriptional control mechanisms underlying flk-1/KDR expression should provide insight into the molecular basis of angiogenesis. In this study, we show that transforming growth factor-beta 1 (TGF-beta 1) down-regulates expression of the endogenous flk-1/KDR gene in endothelial cells. In transient transfection assays, this effect was mapped to a palindromic GATA site in the 5'-untranslated region. In electrophoretic mobility shift assays, the palindromic GATA site was shown to bind to two molecules of GATA protein. Moreover, DNA-GATA interactions were inhibited by TGF-beta 1. Finally, in cotransfection assays, transactivation of the flk-1/KDR promoter by GATA-1 or GATA-2 was attenuated in TGF-beta 1-treated cells. Taken together, these results suggest that the TGF-beta -1-mediated inhibition of the flk-1/KDR gene is mediated by a 5'-untranslated region palindromic GATA site.

* This work was supported by National Institutes of Health Grant P50 HL63609-01 (to W. C. A. and R. D. R.) and by an American Society of Hematology scholar award (to W. C. A.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Biology, 68-480, Massachusetts Institute of Technology, 31 Ames St., Cambridge, MA 02139. Tel.: 617-253-8803; Fax: 617-258-6553; E-mail: rdrrosen@mit.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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