TTBK2 with EB1/3 regulates microtubule dynamics in migrating cells through KIF2A phosphorylation.
TTBK2 with EB1/3 regulates microtubule dynamics in migrating cells through KIF2A phosphorylation.
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DOI：
10.1083/jcb.201412075
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发表时间：
2015-08-31
期刊：
影响因子：
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通讯作者：
Kaibuchi K
中科院分区：
文献类型：
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作者：
Watanabe T;Kakeno M;Matsui T;Sugiyama I;Arimura N;Matsuzawa K;Shirahige A;Ishidate F;Nishioka T;Taya S;Hoshino M;Kaibuchi K
微管（MT）加上末端跟踪蛋白TTBK 2磷酸化驱动蛋白-13家族MT解聚酶KIF 2A，并将其从MT中去除，从而拮抗KIF 2A诱导的MT+末端解聚。微管（MT）在各种细胞事件中发挥关键作用，包括细胞迁移。末端结合蛋白（EBs）在生长的MT末端积累，并通过募集其他+末端跟踪蛋白（+TIPs）来调节MT末端动力学。然而，电子商务如何通过+TIPs促进MT动态仍然是难以捉摸的。我们专注于tau微管蛋白激酶2（TTBK 2）作为EB 1/3结合激酶，并证实TTBK 2作为+TIP。我们确定MT解聚驱动蛋白KIF 2A作为TTBK 2的新底物。TTBK 2在完整细胞中以EB 1/3依赖性方式在S135处磷酸化KIF 2A，并在体外灭活其MT解聚活性。TTBK 2耗竭降低MT寿命（促进收缩和抑制救援）和受损的HeLa细胞迁移，这些表型部分恢复KIF 2A共耗竭。非磷酸化KIF 2A的表达，而不是野生型KIF 2A，减少MT寿命和减缓细胞迁移。这些发现表明，TTBK 2与EB 1/3磷酸化KIF 2A和拮抗KIF 2A诱导的解聚在MT+结束细胞迁移。
The microtubule (MT) plus end–tracking protein TTBK2 phosphorylates kinesin-13 family MT depolymerase KIF2A and removes it from MTs, thereby antagonizing KIF2A-induced depolymerization at MT plus ends during cell migration. Microtubules (MTs) play critical roles in various cellular events, including cell migration. End-binding proteins (EBs) accumulate at the ends of growing MTs and regulate MT end dynamics by recruiting other plus end–tracking proteins (+TIPs). However, how EBs contribute to MT dynamics through +TIPs remains elusive. We focused on tau-tubulin kinase 2 (TTBK2) as an EB1/3-binding kinase and confirmed that TTBK2 acted as a +TIP. We identified MT-depolymerizing kinesin KIF2A as a novel substrate of TTBK2. TTBK2 phosphorylated KIF2A at S135 in intact cells in an EB1/3-dependent fashion and inactivated its MT-depolymerizing activity in vitro. TTBK2 depletion reduced MT lifetime (facilitated shrinkage and suppressed rescue) and impaired HeLa cell migration, and these phenotypes were partially restored by KIF2A co-depletion. Expression of nonphosphorylatable KIF2A, but not wild-type KIF2A, reduced MT lifetime and slowed down the cell migration. These findings indicate that TTBK2 with EB1/3 phosphorylates KIF2A and antagonizes KIF2A-induced depolymerization at MT plus ends for cell migration.