NUCLEAR FACTOR-KAPPA-B INTERACTS FUNCTIONALLY WITH THE PLATELET-DERIVED GROWTH-FACTOR B-CHAIN SHEAR-STRESS RESPONSE ELEMENT IN VASCULAR ENDOTHELIAL-CELLS EXPOSED TO FLUID SHEAR-STRESS
NUCLEAR FACTOR-KAPPA-B INTERACTS FUNCTIONALLY WITH THE PLATELET-DERIVED GROWTH-FACTOR B-CHAIN SHEAR-STRESS RESPONSE ELEMENT IN VASCULAR ENDOTHELIAL-CELLS EXPOSED TO FLUID SHEAR-STRESS
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DOI：
10.1172/jci118106
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发表时间：
1995-08-01
影响因子：
15.9
通讯作者：
COLLINS, T
中科院分区：
文献类型：
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作者：
KHACHIGIAN, LM;RESNICK, N;COLLINS, T
作用于心血管系统内皮细胞的血流动力学力，如流体剪切应力，可以调节越来越多的基因的表达，这些基因对体内平衡和血管疾病的发病机制至关重要，（5 ′-GAGACC-3 ′），以前定义为剪切应力反应元件，存在于许多基因的启动子中，包括PDGF B链，其表达受剪切应力调节，与该元件结合的核蛋白的身份尚未阐明。使用电泳迁移率变动分析和体外DNA酶I足迹法，我们证明了核因子-KB p50-p65异二聚体，其在暴露于流体剪切应力的培养的血管内皮细胞的核中积累，该结合基序的突变消除了其与p50-p65的相互作用，并消除了该启动子介导暴露于剪切应力的内皮细胞中基因表达增加的能力。瞬时共转染研究表明，p50-p65能够激活PDGF-B剪切应力反应元件依赖的报告基因在这些细胞中的表达，这些研究结果，从而牵连核因子-KB的反式激活的内皮基因响应于一个定义的流体机械力。
Hemodynamic forces, such as fluid shear stress, that act on the endothelial lining of the cardiovascular system can modulate the expression of an expanding number of genes crucial for homeostasis and the pathogenesis of vascular disease, A 6-bp core element (5'-GAGACC-3'), defined previously as a shear-stress response element is present in the promoters of many genes, including the PDGF B-chain, whose expression is modulated by shear stress, The identity of the nuclear protein(s) binding to this element has not yet been elucidated, Using electrophoretic mobility shift assays and in vitro DNase I footprinting, we demonstrate that nuclear factor-KB p50-p65 heterodimers, which accumulate in the nuclei of cultured vascular endothelial cells exposed to fluid shear stress, bind to the PDGF-B shear-stress response element in a specific manner, Mutation of this binding motif abrogated its interaction with p50-p65 and abolished the ability of the promoter to mediate increased gene expression in endothelial cells exposed to shear stress. Transient cotransfection studies indicate that p50-p65 is able to activate PDGF-B shear-stress response element-dependent reporter gene expression in these cells, These findings thus implicate nuclear factor-KB in the transactivation of an endothelial gene responding to a defined fluid mechanical force.