Pyk2 regulates cell-edge protrusion dynamics by interacting with Crk.
Pyk2 regulates cell-edge protrusion dynamics by interacting with Crk.
复制标题
DOI：
10.1091/mbc.e20-10-0640
复制
发表时间：
2021-11-01
影响因子：
3.3
通讯作者：
Gil-Henn H
中科院分区：
文献类型：
--
作者：
Lukic N;Lapetina S;Grobe H;Srikanth KD;Twafra S;Solomon J;Sneh T;Gendler M;Zaidel-Bar R;Gil-Henn H
粘着斑激酶 (FAK) 已被广泛认为是细胞迁移的调节因子，但密切相关的富含脯氨酸的酪氨酸激酶 2 (Pyk2) 是否以及如何调节成纤维细胞运动仍存在争议。使用来自 Pyk2–/– 小鼠的小鼠胚胎成纤维细胞 (MEF)，我们在这里首次表明，缺乏 Pyk2 会显着损害随机和定向成纤维细胞运动。 Pyk2–/– MEF 显示细胞边缘突出动力学降低，这取决于 Pyk2 的激酶和蛋白质-蛋白质结合活性。通过体外高通量筛选的生物信息学分析和文本挖掘，我们将 CrkI/II 确定为 Pyk2 的新底物和相互作用子。 CrkI/II 的敲除显示细胞边缘突起的动力学改变，这与 Pyk2–/– MEF 中观察到的表型相似。此外，上位性实验表明，Pyk2 通过与 Crk 的直接和间接相互作用来调节细胞边缘突起的动力学，从而激活和下调 Crk 介导的细胞骨架信号传导。这种复杂的机制一方面可以实现细胞边缘突出动力学和随后的细胞迁移的微调，另一方面可以严格调节细胞运动性，而另一方面，该过程应严格限制在正常细胞的特定时间和环境中。
Focal adhesion kinase (FAK) is well established as a regulator of cell migration, but whether and how the closely related proline-rich tyrosine kinase 2 (Pyk2) regulates fibroblast motility is still under debate. Using mouse embryonic fibroblasts (MEFs) from Pyk2–/– mice, we show here, for the first time, that lack of Pyk2 significantly impairs both random and directed fibroblast motility. Pyk2–/– MEFs show reduced cell-edge protrusion dynamics, which is dependent on both the kinase and protein–protein binding activities of Pyk2. Using bioinformatics analysis of in vitro high- throughput screens followed by text mining, we identified CrkI/II as novel substrates and interactors of Pyk2. Knockdown of CrkI/II shows altered dynamics of cell-edge protrusions, which is similar to the phenotype observed in Pyk2–/– MEFs. Moreover, epistasis experiments suggest that Pyk2 regulates the dynamics of cell-edge protrusions via direct and indirect interactions with Crk that enable both activation and down-regulation of Crk-mediated cytoskeletal signaling. This complex mechanism may enable fine-tuning of cell-edge protrusion dynamics and consequent cell migration on the one hand together with tight regulation of cell motility, a process that should be strictly limited to specific time and context in normal cells, on the other hand.