Negative transcriptional regulation of the VCAM-1 gene by fluid shear stress in murine endothelial cells.
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.
Korenaga
R
R
Department of Cardiovascular Biomechanics, Faculty of Medicine, University of Tokyo, Japan.
Ando
J
J
Kosaki
K
K
Isshiki
M
M
Takada
Y
Y
Kamiya
A
A
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D002478:Cells, Cultured; D016384:Consensus Sequence; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D004727:Endothelium / Q000502:physiology; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D008208:Lymphatic System / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016328:NF-kappa B / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D013314:Stress, Mechanical; D016385:TATA Box; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014158:Transcription, Genetic; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis* / Q000235:genetics*
D017362:Cyclic AMP Response Element-Binding Protein; D016328:NF-kappa B; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1; D019010:Vascular Cell Adhesion Molecule-1