The molecular mechanisms that regulate and coordinate signaling between the extracellular matrix (ECM) and cells contributing to the developing vasculature are complex and poorly understood. Myocardin-like protein 2 (MKL2) is a transcriptional co-activator that in response to RhoA and cytoskeletal actin signals physically associates with serum response factor (SRF), activating a subset of SRF-regulated genes. We now report the discovery of a previously undescribed MKL2/TGFβ signaling pathway in embryonic stem (ES) cells that is required for maturation and stabilization of the embryonic vasculature. Mkl2(-/-) null embryos exhibit profound derangements in the tunica media of select arteries and arterial beds, which leads to aneurysmal dilation, dissection and hemorrhage. Remarkably, TGFβ expression, TGFβ signaling and TGFβ-regulated genes encoding ECM are downregulated in Mkl2(-/-) ES cells and the vasculature of Mkl2(-/-) embryos. The gene encoding TGFβ2, the predominant TGFβ isoform expressed in vascular smooth muscle cells and embryonic vasculature, is activated directly via binding of an MKL2/SRF protein complex to a conserved CArG box in the TGFβ2 promoter. Moreover, Mkl2(-/-) ES cells exhibit derangements in cytoskeletal organization, cell adhesion and expression of ECM that are rescued by forced expression of TGFβ2. Taken together, these data demonstrate that MKL2 regulates a conserved TGF-β signaling pathway that is required for angiogenesis and ultimately embryonic survival.

译文

调节和协调细胞外基质 (ECM) 与有助于发展的脉管系统的细胞之间信号传导的分子机制很复杂,而且知之甚少。Myocardin样蛋白2 (MKL2) 是一种转录共激活剂,可响应RhoA和细胞骨架肌动蛋白信号与血清反应因子 (SRF) 物理相关,从而激活SRF调控基因的一部分。我们现在报告在胚胎干 (ES) 细胞中发现了先前未描述的MKL2/tgf β 信号通路,这是胚胎脉管系统成熟和稳定所必需的。Mkl2(-/-) 无效胚胎在选定的动脉和动脉床的中膜中表现出严重的紊乱,从而导致动脉瘤扩张,解剖和出血。值得注意的是,在Mkl2(-/-) ES细胞和Mkl2(-/-) 胚胎的脉管系统中,tgf β 表达,tgf β 信号传导和编码ECM的tgf β 调节基因被下调。编码TGFβ2 (在血管平滑肌细胞和胚胎脉管系统中表达的主要TGFβ 亚型) 的基因通过将MKL2/SRF蛋白复合物与TGFβ2启动子中的保守的CArG盒结合而直接激活。此外,Mkl2(-/-) ES细胞在细胞骨架组织,细胞粘附和ECM表达方面表现出紊乱,这些紊乱被tgfβ2的强制表达所挽救。总之,这些数据表明,MKL2调节保守的TGF-β 信号通路,这是血管生成和最终胚胎存活所必需的。

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