Requirement of LIM domains for the transient accumulation of paxillin at damaged stress fibres.

Requirement of LIM domains for the transient accumulation of paxillin at damaged stress fibres.
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DOI:
10.1242/bio.20134531
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发表时间:
2013-07-15
期刊:
影响因子:
2.4
通讯作者:
Ikai A
Ikai A
中科院分区:
生物学4区
文献类型:
--
作者:
Watanabe-Nakayama T;Saito M;Machida S;Kishimoto K;Afrin R;Ikai A

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细胞识别并响应细胞内和细胞外机械条件的变化,以维持其机械稳态。肌动蛋白丝和肌球蛋白 II 的线性收缩束(称为应力纤维 (SF))可介导机械信号。机械信号(例如肌球蛋白 II 和/或外力驱动的过度应力)可能会损坏 SF,并诱导 SF 修复复合物(zyxin 和 VASP)在受损部位局部短暂积累。使用安装在荧光显微镜上的原子力显微镜,我们对表达荧光标记的细胞骨架蛋白的细胞施加机械损伤,并记录随后 SF 修复复合物的动员。我们发现 LIM 蛋白桩蛋白 (paxillin) 比 zyxin 更早在受损位点短暂积累,而桩蛋白敲低并不影响 zyxin 易位的动力学。桩蛋白的 C 端一半由四个串联的 LIM 结构域组成,仍然可以易位到 SF 上的受损位点,表明 LIM 结构域对于桩蛋白的机械感觉功能至关重要。我们的研究结果证明了 LIM 结构域在机械传感 LIM 蛋白中的关键作用。
Cells recognize and respond to changes in intra- and extracellular mechanical conditions to maintain their mechanical homeostasis. Linear contractile bundles of actin filaments and myosin II known as stress fibres (SFs) mediate mechanical signals. Mechanical cues such as excessive stress driven by myosin II and/or external force may damage SFs and induce the local transient accumulation of SF-repair complexes (zyxin and VASP) at the damaged sites. Using an atomic force microscope mounted on a fluorescence microscope, we applied mechanical damage to cells expressing fluorescently tagged cytoskeletal proteins and recorded the subsequent mobilization of SF-repair complexes. We found that a LIM protein, paxillin, transiently accumulated at the damaged sites earlier than zyxin, while paxillin knockdown did not affect the kinetics of zyxin translocation. The C-terminal half of paxillin, comprising four-tandem LIM domains, can still translocate to damaged sites on SFs, suggesting that the LIM domain is essential for the mechanosensory function of paxillin. Our findings demonstrate a crucial role of the LIM domain in mechanosensing LIM proteins.
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