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Vol. 273, Issue 5, C1506-C1515, November 1997
1 Department of Cardiovascular
Biomechanics and 2 Institute of
Medical Electronics,
To explore the mechanism of shear stress-induced downregulation
of vascular cell adhesion molecule 1 (VCAM-1) expression in murine
endothelial cells (ECs), we examined the effect of shear stress on
VCAM-1 gene transcription and assessed the
cis-acting elements involved in this
phenomenon. VCAM-1 mRNA expression was downregulated at the
transcriptional level as defined by nuclear run-on assay and transient
transfection of VCAM-1 promoter-luciferase gene constructs. The
luciferase assay on the VCAM-1 deletion mutants revealed that the
cis-acting element is contained
between 
694 and 329 bp upstream from the transcription
initiation site. Gel shift assay using overlapping oligonucleotide
probes of this region showed that oligonucleotides containing a double
AP-1 consensus sequence (TGACTCA) formed distinct complexes with
nuclear proteins extracted from shear-stressed cells. Mutation of
either one or both of two AP-1 consensus sequences completely abolished
the ability of the promoter to respond to shear stress. These results suggest that fluid shear stress downregulates the transcription of the
VCAM-1 gene via an upstream
cis-element, a double AP-1 consensus
sequence, in murine lymph node venule ECs.
vascular endothelial cells; vascular cell adhesion molecule 1; nuclear factor activator protein-1; c-jun
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