Yinchen Dong, Ahmed Elgerbi, Bin Xie, John S Choy, Sanjeevi Sivasankar
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引用次数: 0
Abstract
Desmosomes are essential cell-cell adhesion organelles that enable tension-prone tissue, like the skin and heart, to withstand mechanical stress. Desmosomal anomalies are associated with numerous epidermal disorders and cardiomyopathies. Despite their critical role in maintaining tissue resilience, an understanding of how desmosomes sense and respond to mechanical stimuli is lacking. Here, we use a combination of super-resolution imaging, FRET-based tension sensors, atomistic computer simulations, and biochemical assays to demonstrate that actomyosin forces induce a conformational change in desmoplakin, a critical cytoplasmic desmosomal protein. We show that in human breast cancer MCF7 cells, actomyosin contractility reorients keratin intermediate filaments and directs force to desmoplakin along the keratin filament backbone. These forces induce a conformational change in the N-terminal plakin domain of desmoplakin, converting this domain from a folded (closed) to an extended (open) conformation. Our findings establish that desmoplakin is mechanosensitive and responds to changes in cellular load by undergoing a force-induced conformational change.
脱模小体是细胞-细胞粘附的重要细胞器,能使皮肤和心脏等易受张力影响的组织承受机械压力。脱丝体异常与许多表皮疾病和心肌病有关。尽管脱丝体在维持组织韧性方面起着至关重要的作用,但人们对脱丝体如何对机械刺激做出反应还缺乏了解。在这里,我们在人类乳腺癌 MCF7 细胞中证明,肌动蛋白的作用力会诱导脱丝体蛋白 N 端区域的构象变化,脱丝体蛋白是一种关键的细胞质脱丝体蛋白。利用超分辨率荧光显微镜、生化试验和原子计算机模拟,我们发现肌动蛋白的作用力是沿着角蛋白-19细丝导向去角质蛋白的。这些作用力诱导去鳞屑蛋白 N 端 plakin 结构域发生构象变化,使该结构域从折叠(封闭)构象转化为扩展(开放)构象。功能性粘附试验表明,与处于封闭状态的细胞相比,处于开放构象的去瘤素MCF7细胞更具粘附性。我们的研究结果证实,去鳞屑蛋白对机械敏感,会发生由力诱导的构象变化,从而增强去鳞屑体的机械弹性:脱丝体是重要的细胞-细胞粘附连接体,可使皮肤和心脏等易受张力影响的组织承受机械应力。尽管它们在维持组织完整性方面发挥着关键作用,但目前尚不清楚脱模小体如何对机械力做出反应。在这里,我们证明了作用力会诱导细胞内的一种重要脱膜体蛋白--脱膜蛋白发生构象变化。这种由力诱导的脱膜蛋白结构变化增强了脱膜体的复原力,使它们能够承受机械应力。