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Papers In Press, published online ahead of print April 8, 2004
Cardiology, Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02215
Corresponding Author: JLi{at}BIDMC.Harvard.edu
VEGF is an angiogenic growth factor known to be upregulated in ischemic heart and hypoxic endothelial cells. However, the transcriptional regulation of VEGF in hypoxia-induced angiogenesis is not fully understood. TEF-1 is a transcriptional factor family that can regulate many genes expressed in cardiac and skeletal muscle cells by binding to M-CAT elements in the promoters of these genes. In this study, we demonstrated that Related Transcriptional Enhance Factor-1 (RTEF-1), a member of the TEF-1 family, is upregulated in hypoxic endothelial cells. Over-expression of RTEF-1 increases VEGF promoter activity and VEGF expression. Sequential deletion and site-directed mutation analyses of the VEGF promoter demonstrated that a GC-rich region containing four Sp1 response elements, located between –114 and –50, was essential for RTEF-1 function. This region is beyond of HIF-1a binding site and does not consist of M-CAT related elements. By electrophoretic mobility shift assay, RTEF-1 was found to interact with the first Sp1 residue (–97 ~ –87) of the four consecutive Sp1 elements. Binding activity of RTEF-1 to VEGF promoter is also confirmed by chromatin immunoprecipitation. In addition, induction of VEGF promoter activity by RTEF-1 results in an increase of angiogeneic processes including endothelial cells proliferation and vascular structure formation. These results indicate that RTEF-1 acts as a transcriptional stimulator of VEGF by regulating VEGF promoter activity through binding to Sp1 site. In addition, RTEF-1-induced VEGF promoter activity was enhanced in a hypoxic condition, indicating that RTEF-1 may play an important role in regulation of VEGF under hypoxia.
J. Biol. Chem, 10.1074/jbc.M403103200
Submitted on March 19, 2004
Revised on April 8, 2004
Accepted on April 8, 2004
RTEF-1, a novel transcriptional stimulator of VEGF in hypoxic endothelial cells
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