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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2003;23:2041.)
© 2003 American Heart Association, Inc.

Vascular Biology

Ets Motifs Are Necessary for Endothelial Cell–Specific Expression of a 723-bp Tie-2 Promoter/Enhancer in Hprt Targeted Transgenic Mice

Takashi Minami; Jan Albert Kuivenhoven; Valerie Evans; Tatsuhiko Kodama; Robert D. Rosenberg; William C. Aird

From the Research Center for Advanced Science and Technology (T.M., T.K.), University of Tokyo, Japan; Department of Molecular Medicine (T.M., W.C.A., R.D.R.), Beth Israel Deaconess Medical Center/Harvard Medical School, Boston, Mass; and Department of Biology (J.A.K., V.E., R.D.R.), Massachusetts Institute of Technology, Cambridge, Mass.

Correspondence to Takashi Minami, Research Center for Advanced Science and Technology, University of Tokyo, 4-6-1 Komaba, Meguro Tokyo, 153-8904. E-mail minami{at}med.rcast.u-tokyo.ac.jp

Objective— Tie-2 is an endothelial cell–specific receptor tyrosine kinase that is involved in the remodeling of blood vessels and angiogenesis. Our goal was to characterize Tie-2 promoter function as a means of providing insight into the mechanisms of endothelial cell–specific gene regulation.

Methods and Results— When targeted to the Hprt locus of mice, a small Tie-2 promoter fragment (containing a 300-bp intronic enhancer coupled upstream to a 423-bp core promoter) (T-short) directed widespread endothelial cell expression in vivo. The T-short promoter contains 2 clusters of Ets sites, one in the first exon, the other in the intronic enhancer. In cultured endothelial cells, a combined mutation of the Ets motifs resulted in a significant reduction in promoter activity. Consistent with these results, the same Ets mutations resulted in a loss of detectable expression of the T-short promoter in all vascular beds with the notable exception of the brain.

Conclusions— These results suggest that the T-short promoter contains information for widespread expression in the vascular tree, Ets sites are necessary for in vivo promoter activity, and the shorter Tie-2 fragment may be useful as a tool to direct heterologous gene expression within the intact endothelium.

Key Words: Tie-2 • endothelial cells • transgenic mice • gene regulation • Ets motifs

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