NudCL2 regulates cell migration by stabilizing both myosin-9 and LIS1 with Hsp90
NudCL2 regulates cell migration by stabilizing both myosin-9 and LIS1 with Hsp90
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NudCL2 通过用 Hsp90 稳定肌球蛋白 9 和 LIS1 来调节细胞迁移
DOI：
10.1038/s41419-020-02739-9
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发表时间：
2020-07-14
影响因子：
9
通讯作者：
Yang, Yuehong
中科院分区：
文献类型：
--
作者：
Chen, Wenwen;Wang, Wei;Yang, Yuehong
细胞迁移在许多生物学过程中起着关键作用，但其机制尚不清楚。在这里，我们发现NudC样蛋白2（NudCL 2），热休克蛋白90（Hsp 90）的辅助伴侣，调节细胞迁移稳定肌球蛋白9和无脑蛋白1（LIS 1）。敲除或敲除NudCL 2显著增加单细胞迁移，但对集体细胞迁移没有显著影响。免疫沉淀-质谱和蛋白质印迹分析表明，NudCL 2结合肌球蛋白-9在哺乳动物细胞。NudCL 2的耗尽不仅降低肌球蛋白-9蛋白水平，而且导致肌动蛋白解体。肌球蛋白-9的异位表达有效地逆转了NudCL 2缺失细胞中肌动蛋白解体和单细胞迁移的缺陷。有趣的是，肌球蛋白-9的敲低增加单个和集体细胞迁移。NudCL 2客户蛋白LIS 1的耗尽抑制单个和集体细胞迁移，这与肌球蛋白9耗尽相比表现出相反的效果。肌球蛋白-9和LIS 1的共耗竭促进单细胞迁移，类似于NudCL 2耗竭引起的表型。此外，Hsp 90 ATP酶活性的抑制也降低了Hsp 90相互作用蛋白肌球蛋白-9的稳定性，并增加单细胞迁移。Hsp 90的强制表达有效地逆转肌球蛋白-9蛋白的不稳定性和由NudCL 2耗尽诱导的缺陷，但反之亦然。总之，这些数据表明，NudCL 2通过Hsp 90途径稳定肌球蛋白-9和LIS 1在精确调节细胞迁移中起重要作用。
Cell migration plays pivotal roles in many biological processes; however, its underlying mechanism remains unclear. Here, we find that NudC-like protein 2 (NudCL2), a cochaperone of heat shock protein 90 (Hsp90), modulates cell migration by stabilizing both myosin-9 and lissencephaly protein 1 (LIS1). Either knockdown or knockout of NudCL2 significantly increases single-cell migration, but has no significant effect on collective cell migration. Immunoprecipitation–mass spectrometry and western blotting analyses reveal that NudCL2 binds to myosin-9 in mammalian cells. Depletion of NudCL2 not only decreases myosin-9 protein levels, but also results in actin disorganization. Ectopic expression of myosin-9 efficiently reverses defects in actin disorganization and single-cell migration in cells depleted of NudCL2. Interestingly, knockdown of myosin-9 increases both single and collective cell migration. Depletion of LIS1, a NudCL2 client protein, suppresses both single and collective cell migration, which exhibits the opposite effect compared with myosin-9 depletion. Co-depletion of myosin-9 and LIS1 promotes single-cell migration, resembling the phenotype caused by NudCL2 depletion. Furthermore, inhibition of Hsp90 ATPase activity also reduces the Hsp90-interacting protein myosin-9 stability and increases single-cell migration. Forced expression of Hsp90 efficiently reverses myosin-9 protein instability and the defects induced by NudCL2 depletion, but not vice versa. Taken together, these data suggest that NudCL2 plays an important role in the precise regulation of cell migration by stabilizing both myosin-9 and LIS1 via Hsp90 pathway.