Focal adhesion kinase is involved in mechanosensing during fibroblast migration
Focal adhesion kinase is involved in mechanosensing during fibroblast migration
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DOI：
10.1073/pnas.201201198
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发表时间：
2001-09-25
影响因子：
11.1
通讯作者：
Wang, YL
中科院分区：
文献类型：
--
作者：
Wang, HB;Dembo, M;Wang, YL
粘着斑激酶（FAK）是一种位于粘着斑的非受体蛋白酪氨酸激酶，被认为介导了粘着刺激效应。虽然FAK的消融损害细胞运动，但尚不清楚FAK是否可能参与细胞迁移的指导，这一作用与其假定的调节功能一致。在四环素抑制系统的控制下，我们用FAK基因转染了FAK-null成纤维细胞。将细胞培养在柔性聚丙烯酰胺基质上，用于检测牵引力和施加机械刺激。与表达野生型FAK的对照细胞相比，FAK缺失细胞的迁移速度和方向持久性降低。此外，表达FAK的细胞通过重新定向其运动并形成明显的局灶性粘连来对施加的力做出反应，而无FAK的细胞则未能表现出此类反应。此外，FAK-空细胞表现出受损的反应，降低基板的灵活性，这导致控制细胞产生较弱的牵引力和迁移远离软基板。细胞表达Y397 F FAK，不能在一个关键的酪氨酸位点磷酸化，表现出类似的缺陷，迁移模式和力诱导的重定向没有FAK-空细胞。然而，F397-FAK细胞的其他方面，包括对底物柔性的反应和机械刺激后粘着斑的扩增，与对照细胞相似。我们的研究结果表明，FAK在迁移细胞对机械输入的反应中起着重要作用。此外，Tyr-397的磷酸化是FAK在细胞迁移中的一些但不是全部功能所必需的。
Focal adhesion kinase (FAK) is a non-receptor protein tyrosine kinase localized at focal adhesions and is believed to mediate adhesion-stimulated effects. Although ablation of FAK impairs cell movement, it is not clear whether FAK might be involved in the guidance of cell migration, a role consistent with its putative regulatory function. We have transfected FAK-null fibroblasts with FAK gene under the control of the tetracycline repression system. Cells were cultured on flexible polyacrylamide substrates for the detection of traction forces and the application of mechanical stimulation. Compared with control cells expressing wild-type FAK, FAK-null cells showed a decrease in migration speed and directional persistence. In addition, whereas FAK-expressing cells responded to exerted forces by reorienting their movements and forming prominent focal adhesions, FAK-null cells failed to show such responses. Furthermore, FAK-null cells showed impaired responses to decreases in substrate flexibility, which causes control cells to generate weaker traction forces and migrate away from soft substrates. Cells expressing Y397F FAK, which cannot be phosphorylated at a key tyrosine site, showed similar defects in migration pattern and force-induced reorientation as did FAK-null cells. However, other aspects of F397-FAK cells, including the responses to substrate flexibility and the amplification of focal adhesions upon mechanical stimulation, were similar to that of control cells. Our results suggest that FAK plays an important role in the response of migrating cells to mechanical input. In addition, phosphorylation at Tyr-397 is required for some, but not all, of the functions of FAK in cell migration.