Focal Adhesion Kinase Stabilizes the Cytoskeleton
Focal Adhesion Kinase Stabilizes the Cytoskeleton
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DOI：
10.1016/j.bpj.2011.09.043
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发表时间：
2011-11-02
影响因子：
3.4
通讯作者：
Goldmann, Wolfgang H.
中科院分区：
文献类型：
--
作者：
Fabry, Ben;Klemm, Anna H.;Goldmann, Wolfgang H.
粘着斑激酶（FAK）是一种促进粘着斑转换的中心粘着斑蛋白，但FAK对细胞机械稳定性的作用尚不清楚。我们测量了机械性能的野生型（FAKwt），FAK缺陷（FAK-/-），FAK沉默（siFAK），和siControl小鼠胚胎成纤维细胞的磁镊子，原子力显微镜，牵引显微镜，和纳米粒子跟踪微流变学。与野生型细胞相比，FAK缺陷型细胞表现出较低的细胞硬度，降低的粘附强度和增加的细胞骨架动力学。这些观察结果意味着在FAK缺陷细胞中细胞骨架的稳定性降低。我们将降低的细胞骨架稳定性归因于FAK缺陷细胞中rho激酶的激活，其抑制有序应力纤维束的形成，增强皮质肌动蛋白分布，并减少细胞扩散。与这种解释一致的是，在用Y27632抑制rho激酶后，FAK-/-细胞中的细胞硬度和细胞骨架稳定性部分恢复到野生型水平。
Focal adhesion kinase (FAK) is a central focal adhesion protein that promotes focal adhesion turnover, but the role of FAK for cell mechanical stability is unknown. We measured the mechanical properties of wild-type (FAKwt), FAK-deficient (FAK-/-), FAK-silenced (siFAK), and siControl mouse embryonic fibroblasts by magnetic tweezer, atomic force microscopy, traction microscopy, and nanoscale particle tracking microrheology. FAK-deficient cells showed lower cell stiffness, reduced adhesion strength, and increased cytoskeletal dynamics compared to wild-type cells. These observations imply a reduced stability of the cytoskeleton in FAK-deficient cells. We attribute the reduced cytoskeletal stability to rho-kinase activation in FAK-deficient cells that suppresses the formation of ordered stress fiber bundles, enhances cortical actin distribution, and reduces cell spreading. In agreement with this interpretation is that cell stiffness and cytoskeletal stability in FAK-/- cells is partially restored to wild-type level after rho-kinase inhibition with Y27632.