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J Biol Chem, Vol. 274, Issue 29, 20376-20383, July 16, 1999
From the The TIE2 gene, also known as
TEK, encodes a tyrosine kinase receptor that is required
for the normal development of the vascular system during embryogenesis.
TIE2 is specifically expressed in endothelial cells;
however, the transcriptional mechanisms that regulate this highly
restricted pattern of expression remain unknown. Here we demonstrate
that a consensus octamer element located in the 5'-flanking region of
TIE2 is required for normal expression in embryonic
endothelial cells. Transgenic embryos carrying a TIE2/LacZ
construct spanning 2.1 kilobases of upstream regulatory sequences
exhibit expression of the reporter transgene specifically in
endothelial cells. Site-directed mutagenesis of a consensus octamer
element located in this region results in the loss of enhancer
activity and significantly impairs the endothelial expression of
the reporter transgene. Consistent with the in vivo
data, in vitro DNA-protein binding studies show that the
consensus octamer element displays an endothelial cell-specific pattern
of binding, suggesting an interaction with a protein complex consisting
of Oct1 and an endothelial cell-restricted cofactor. These data
identify a novel role for the octamer element as an essential regulator of TIE2 expression, define the first known transcriptional
pathway that mediates the expression of a developmental endothelial
cell gene, and provide insights into the transcriptional mechanisms that regulate development of the vasculature during embryogenesis.
Octamer-dependent in Vivo Expression of
the Endothelial Cell-specific TIE2 Gene
§,§, and
Division of Cardiovascular Medicine, Falk
Cardiovascular Research Center, Stanford University, Stanford,
California 94305-5406 and the § Cardiology Section, Palo
Alto Veterans Affairs Medical Center,
Palo Alto, California 94304
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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