Arrestins regulate cell spreading and motility via focal adhesion dynamics.
Arrestins regulate cell spreading and motility via focal adhesion dynamics.
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DOI：
10.1091/mbc.e14-02-0740
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发表时间：
2015-02-15
影响因子：
3.3
通讯作者：
Gurevich VV
中科院分区：
文献类型：
--
作者：
Cleghorn WM;Branch KM;Kook S;Arnette C;Bulus N;Zent R;Kaverina I;Gurevich EV;Weaver AM;Gurevich VV
缺乏这两种非视觉抑制蛋白的细胞表现出过度扩散、粘着斑解体缺陷和对微管敏感。这种表型是拯救野生型arrestins，但不是突变体缺乏网格蛋白结合，这表明arrestins连接微管和网格蛋白调节局灶性粘附拆卸。粘着斑（FAs）在细胞粘附中起关键作用，并且它们的及时拆卸是细胞运动所必需的。微管依赖性靶向和网格蛋白的募集对于FA分解都是至关重要的。在这里，我们确定非视觉arrestins之间的微管和网格蛋白的分子联系。缺乏这两种非视觉抑制蛋白的细胞表现出在纤连蛋白和聚d-赖氨酸上过度扩散、粘附增加和运动性降低。抑制蛋白的缺乏大大增加了脂肪酸的大小和寿命，表明抑制蛋白是快速脂肪酸周转所必需的。在诺考达唑洗脱试验中，arrestin缺陷细胞中的FA对微管的解离或再生长无反应，这表明arrestins对微管靶向依赖性FA解离是必要的。网格蛋白在抑制蛋白缺陷细胞中FA附近表现出动力学降低。与野生型arrestins相反，缺乏网格蛋白结合的突变体没有拯救表型。总的来说，这些数据表明，抑制蛋白是连接微管和网格蛋白的FA分解的关键调节因子。
Cells lacking both nonvisual arrestins show excessive spreading, defects in focal adhesion disassembly, and sensitivity to microtubules. This phenotype is rescued by wild-type arrestins but not mutants deficient in clathrin binding, suggesting that arrestins regulate focal adhesion disassembly by linking microtubules and clathrin. Focal adhesions (FAs) play a key role in cell attachment, and their timely disassembly is required for cell motility. Both microtubule-dependent targeting and recruitment of clathrin are critical for FA disassembly. Here we identify nonvisual arrestins as molecular links between microtubules and clathrin. Cells lacking both nonvisual arrestins showed excessive spreading on fibronectin and poly-d-lysine, increased adhesion, and reduced motility. The absence of arrestins greatly increases the size and lifespan of FAs, indicating that arrestins are necessary for rapid FA turnover. In nocodazole washout assays, FAs in arrestin-deficient cells were unresponsive to disassociation or regrowth of microtubules, suggesting that arrestins are necessary for microtubule targeting–dependent FA disassembly. Clathrin exhibited decreased dynamics near FA in arrestin-deficient cells. In contrast to wild-type arrestins, mutants deficient in clathrin binding did not rescue the phenotype. Collectively the data indicate that arrestins are key regulators of FA disassembly linking microtubules and clathrin.