Multiple cis-elements mediate shear stress-induced gene expression.
Fluid shear stress activates the expression of immediate early (IE) genes in vascular endothelial cells. The transcriptional regulation can be mediated through the shear stress-sensitive cis-acting elements at the 5' promoter regions of various IE genes such as the monocyte chemotactic protein-1 (MCP-1) gene. We linked wild-type and mutated MCP-1 promoters to the reporter gene luciferase and used such constructs to investigate the role of the phorbol ester TPA responsive element (TRE) in the shear-induced MCP-1 gene expression in vascular endothelial cells. Functional analysis showed that TGACTCC (a divergent TRE) located at nt -54 to -60 is necessary for shear-inducibility in bovine aortic endothelial cells (BAEC). The induction of the wild-type MCP-1 promoter construct by shear stress was attenuated by pretreating the cells with 1 microM dexamethasone or 1 microM retinoic acid 12 h before the shear stress experiments. The induction by shear stress reduced from 13-fold in the untreated cells to 7- and 3-folds in the dexamethasone- and retinoic acid-treated cells, respectively. These results demonstrate that the glucocorticoid receptor and retinoic acid receptor may interfere with the shear stress-activated AP-1/TRE. The reporter activity of HIV(LTR), which is a plasmid construct of the long terminal repeats of the human immunodeficiency virus and contains a kappa B enhancer element, was also activated by shear stress. The results of our investigations indicate that the shear stress-induced IE gene expression can be mediated through multiple cis-elements.
Shyy
J Y
JY
Department of Bioengineering, University of California, La Jolla 92093-0412, USA. Schien@bioeng.ucsd.edu.
Li
Y S
YS
Lin
M C
MC
Chen
W
W
Yuan
S
S
Usami
S
S
Chien
S
S
D000818:Animals; D002273:Carcinogens / Q000494:pharmacology; D002417:Cattle; D018932:Chemokine CCL2 / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D017781:Genes, Immediate-Early / Q000187:drug effects / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D015752:Genes, env / Q000235:genetics; D005938:Glucocorticoids / Q000494:pharmacology; D007641:Keratolytic Agents / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D011965:Receptors, Glucocorticoid / Q000235:genetics; D018168:Receptors, Retinoic Acid / Q000235:genetics; D012212:Rheology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D014212:Tretinoin / Q000494:pharmacology
D002273:Carcinogens; D018932:Chemokine CCL2; D005938:Glucocorticoids; D007641:Keratolytic Agents; D011965:Receptors, Glucocorticoid; D018168:Receptors, Retinoic Acid; D018808:Transcription Factor AP-1; D014212:Tretinoin; D003907:Dexamethasone; D008156:Luciferases; D013755:Tetradecanoylphorbol Acetate