Electron Microscope Laboratory, RIKEN Center for Developmental Biology, Kobe, Hyogo 650-0047, Japan.
Nat Cell Biol. 2010 Jun;12(6):533-42. doi: 10.1038/ncb2055. Epub 2010 May 9.
Adherens junctions (AJs), which are organized by adhesion proteins and the underlying actin cytoskeleton, probably sense pulling forces from adjacent cells and modulate opposing forces to maintain tissue integrity, but the regulatory mechanism remains unknown at the molecular level. Although the possibility that alpha-catenin acts as a direct linker between the membrane and the actin cytoskeleton for AJ formation and function has been minimized, here we show that alpha-catenin recruits vinculin, another main actin-binding protein of AJs, through force-dependent changes in alpha-catenin conformation. We identified regions in the alpha-catenin molecule that are required for its force-dependent binding of vinculin by introducing mutant alpha-catenin into cells and using in vitro binding assays. Fluorescence recovery after photobleaching analysis for alpha-catenin mobility and the existence of an antibody recognizing alpha-catenin in a force-dependent manner further supported the notion that alpha-catenin is a tension transducer that translates mechanical stimuli into a chemical response, resulting in AJ development.
黏着连接(AJs)由黏附蛋白和下方的肌动蛋白细胞骨架组成,可能感知来自相邻细胞的拉力,并调节反作用力以维持组织完整性,但分子水平上的调控机制仍不清楚。尽管α-连环蛋白作为 AJ 形成和功能的膜和肌动蛋白细胞骨架之间的直接连接物的可能性已经最小化,但在这里我们表明,α-连环蛋白通过α-连环蛋白构象的力依赖性变化募集粘着斑蛋白,另一种 AJ 的主要肌动蛋白结合蛋白。我们通过将突变型α-连环蛋白引入细胞并使用体外结合测定法,鉴定了α-连环蛋白分子中需要其力依赖性结合粘着斑蛋白的区域。荧光恢复后光漂白分析α-连环蛋白的迁移率以及以力依赖性方式识别α-连环蛋白的抗体的存在进一步支持了这样的观点,即α-连环蛋白是一种张力转导蛋白,可将机械刺激转化为化学反应,从而导致 AJ 的发育。