Phosphorylation of cortactin by cyclin-dependent kinase 5 modulates actin bundling by the dynamin 1-cortactin ring-like complex and formation of filopodia and lamellipodia in NG108-15 glioma-derived cells
Phosphorylation of cortactin by cyclin-dependent kinase 5 modulates actin bundling by the dynamin 1-cortactin ring-like complex and formation of filopodia and lamellipodia in NG108-15 glioma-derived cells
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DOI：
10.3892/ijo.2018.4663
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发表时间：
2019-02-01
影响因子：
5.2
通讯作者：
Yamada, Hiroshi
中科院分区：
文献类型：
--
作者：
Abe, Tadashi;La, The Mon;Yamada, Hiroshi
发动蛋白与皮质蛋白共聚形成环状复合物，使肌动蛋白丝成束并稳定。肌动蛋白束的形成对于丝状伪足和板状伪足的产生至关重要，丝状伪足和板状伪足引导癌细胞的迁移、侵袭和转移。然而，它是未知的动力蛋白-coronin复合物如何调节肌动蛋白束的形成。本研究探讨了细胞周期蛋白依赖性激酶5（CDK 5）的磷酸化coronin及其对肌动蛋白束形成的动力蛋白coronin复合物的影响。CDK 5在体外直接在T145/T219处磷酸化coronin。使用其中这些苏氨酸残基中的一个或两个被天冬氨酸取代的拟磷酸突变体。三个磷酸化模拟突变体（T145 D、T219 D和T145 DT 219 D）对F-肌动蛋白的亲和力降低。此外，电子显微镜表明，这些磷酸模拟突变体不能形成一个环状复合物与发动蛋白1。一致，动力蛋白1-磷酸模拟corneumin复合物表现出减少肌动蛋白捆绑活动。拟磷酸化突变体的表达不仅导致异常板状伪足和短丝状伪足，而且导致NG 108 -15胶质瘤衍生细胞的细胞迁移。这些结果表明，磷酸化的corpine由CDK 5调节板状伪足和丝状伪足的形成，通过调节发动蛋白1/皮质素依赖的肌动蛋白捆绑。总之，这些发现表明CDK 5是抗癌治疗的潜在分子靶点。
Dynamin copolymerizes with cortactin to form a ring-like complex that bundles and stabilizes actin filaments. Actin bundle formation is crucial for generation of filopodia and lamellipodia, which guide migration, invasion, and metastasis of cancer cells. However, it is unknown how the dynamin-cortactin complex regulates actin bundle formation. The present study investigated phosphorylation of cortactin by cyclin-dependent kinase 5 (CDK5) and its effect on actin bundle formation by the dynamin-cortactin complex. CDK5 directly phosphorylated cortactin at T145/T219 in vitro. Phosphomimetic mutants in which one or both of these threonine residues was substituted by aspartate were used. The three phosphomimetic mutants (T145D, T219D and T145DT219D) had a decreased affinity for F-actin. Furthermore, electron microscopy demonstrated that these phosphomimetic mutants could not form a ring-like complex with dynamin 1. Consistently, the dynamin 1-phosphomimetic cortactin complexes exhibited decreased actin-bundling activity. Expression of the phosphomimetic mutants resulted in not only aberrant lamellipodia and short filopodia but also cell migration in NG108-15 glioma-derived cells. These results indicate that phosphorylation of cortactin by CDK5 regulates formation of lamellipodia and filopodia by modulating dynamin 1/cortactin-dependent actin bundling. Taken together, these findings suggest that CDK5 is a potential molecular target for anticancer therapy.