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J. Biol. Chem., Vol. 277, Issue 28, 25143-25151, July 12, 2002

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Ets-1 Regulates fli-1 Expression in Endothelial Cells
IDENTIFICATION OF ETS BINDING SITES IN THE fli-1 GENE PROMOTER^*

Etienne Lelièvre, Frédéric Lionneton, Virginie Mattot, Nathalie Spruyt, and Fabrice Soncin

From CNRS UMR 8526, Institut de Biologie de Lille, 1 rue Calmette, 59021 Lille Cedex, France

To understand the role of the Ets-1 transcription factor during angiogenesis, we have overexpressed it in endothelial cells and analyzed the levels of expression of several candidate target genes involved in angiogenesis. The transcripts levels of the ETS transcription factor fli-1 are specifically up-regulated in endothelial cells, which overexpress Ets-1, but not in fibroblasts. Analysis of the promoter of the mouse fli-1 gene reveals that the 1-kb region that comprises the transcription starts and part of exon 1 is responsible for the response of the promoter to Ets-1. The 270/41 fragment contains two known Spi-1-responding Ets binding sites (EBS), which are also necessary for the activation by Ets-1. In contrast to Spi-1, a third EBS is necessary for the full response of this promoter fragment to Ets-1. The rest of the promoter activity has been located in the 986/505 region, where three active EBSs have been identified. Furthermore, endogenous Fli-1 was found to be bound to its own gene promoter and to be able to promote the transactivation of its gene. These results suggest that Ets-1 activates an auto-regulatory loop of expression of fli-1 in endothelial cells, a mechanism that could have significant implications for the endothelial cell fate.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF483910.

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