Regulation of keratin 5/14 intermediate filaments by CDK1, Aurora-B, and Rho-kinase
Regulation of keratin 5/14 intermediate filaments by CDK1, Aurora-B, and Rho-kinase
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CDK1、Aurora-B 和 Rho 激酶对角蛋白 5/14 中间丝的调节
DOI：
10.1016/j.bbrc.2018.03.016
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Inagaki M
中科院分区：
文献类型：
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作者：
Inaba H;Yamakawa D;Tomono Y;Enomoto A;Mii S;Kasahara K;Goto H;Inagaki M
我们以前报道，波形蛋白，GFAP，结蛋白（III型中间丝[IF]蛋白）的有丝分裂磷酸化的CDK 1，极光-B，和Rho激酶。这种磷酸化对于这些IF的有效分离和胞质分裂的完成是至关重要的。角蛋白5（K5）与K14形成异源二聚体，在复层鳞状上皮基底层细胞中构成IF网络。在这里，我们报告，K5/K14的溶解度增加有丝分裂。体外试验表明，三种有丝分裂激酶磷酸化K5的程度高于K14。然后，我们确定Thr 23/Thr 144，Ser 30，和Thr 159对小鼠K5作为主要的磷酸化位点的CDK 1，极光-B，和Rho激酶，分别。使用网站和磷酸化状态特异性抗体，我们证明，K5-Thr 23磷酸化在整个细胞质中从中期到中期，而K5-Ser 30磷酸化发生在分裂沟从后期到末期。有效的K5/K14-IF分离受损的K5突变的网站磷酸化这些有丝分裂激酶。K5-Thr 23磷酸化在小鼠个体的分裂K5阳性细胞中广泛检测到。这些结果表明，K5/K14-IF网络的有丝分裂重组在很大程度上是通过CDK 1，Aurora-B和Rho激酶的K5磷酸化来控制的。
We previously reported that vimentin, GFAP, and desmin (type III intermediate filament [IF] proteins) are mitotically phosphorylated by CDK1, Aurora-B, and Rho-kinase. This phosphorylation is critical for efficient separation of these IFs and completion of cytokinesis. Keratin 5 (K5) and K14 form a heterodimer, which constitutes IF network in basal layer cells of stratified squamous epithelia. Here, we report that the solubility of K5/K14 increased in mitosis. Thein vitroassays revealed that three mitotic kinases phosphorylate K5 more than K14. We then identified Thr23/Thr144, Ser30, and Thr159 on murine K5 as major phosphorylation sites for CDK1, Aurora-B, and Rho-kinase, respectively. Using site- and phosphorylation-state-specific antibodies, we demonstrated that K5-Thr23 was phosphorylated in entire cytoplasm from prometaphase to metaphase, whereas K5-Ser30 phosphorylation occurred specifically at the cleavage furrow from anaphase to telophase. Efficient K5/K14-IF separation was impaired by K5 mutations at the sites phosphorylated by these mitotic kinases. K5-Thr23 phosphorylation was widely detected in dividing K5-positive cells of murine individuals. These results suggested that mitotic reorganization of K5/K14-IF network is governed largely through K5 phosphorylation by CDK1, Aurora-B, and Rho-kinase.