Clathrin heavy chain phosphorylated at T606 plays a role in proper cell division
Clathrin heavy chain phosphorylated at T606 plays a role in proper cell division
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DOI：
10.1080/15384101.2019.1637201
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发表时间：
2019-07
期刊：
影响因子：
4.3
通讯作者：
Yusuke Yabuno;T. Uchihashi;Towa Sasakura;Hiroyuki Shimizu;Y. Naito;K. Fukushima;Kaori Ota;M. Kogo;H. Nojima;N. Yabuta
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文献类型：
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作者：
Yusuke Yabuno;T. Uchihashi;Towa Sasakura;Hiroyuki Shimizu;Y. Naito;K. Fukushima;Kaori Ota;M. Kogo;H. Nojima;N. Yabuta
摘要胞膜蛋白除了调节细胞膜转运外，还调节有丝分裂进程。然而，在有丝分裂过程中，笼蛋白的详细调控机制仍然不清楚。在这里，我们展示了在细胞周期有丝分裂阶段对笼状蛋白的新的调节。网状蛋白重链(Chathrin Heavy Chain，CHC)在T606位被其结合伙伴细胞周期蛋白G相关蛋白(Cyclin G-Associated Kinase，GAK)磷酸化。这种磷酸化对于移植到裸鼠体内的细胞的正常增殖和肿瘤生长是必需的。免疫荧光分析表明，CHC-pT606信号在有丝分裂过程中的定位发生了变化。间期CHC-pT606信号定位于细胞核和中心体，而CHC信号主要分布在细胞质。免疫共沉淀表明CHC与GAK和Polo-like kinase1(PLK1)形成了复合体。使用siRNA耗尽GAK导致中期停滞和CHC-pT606的异常定位，从而消除中期染色质上的KiZ-pT379(作为PLK1的磷酸化靶标)信号。综上所述，我们认为GAK_CHC-pT606_PLK1_KiZ-pT379轴在癌细胞的增殖中起作用。
ABSTRACT Clathrin regulates mitotic progression, in addition to membrane trafficking. However, the detailed regulatory mechanisms of clathrin during mitosis remain elusive. Here, we demonstrate novel regulation of clathrin during mitotic phase of the cell cycle. Clathrin heavy chain (CHC) was phosphorylated at T606 by its association partner cyclin G-associated kinase (GAK). This phosphorylation was required for proper cell proliferation and tumor growth of cells implanted into nude mice. Immunofluorescence analysis showed that the localization of CHC-pT606 signals changed during mitosis. CHC-pT606 signals localized in the nucleus and at the centrosome during interphase, whereas CHC signals were mostly cytoplasmic. Co-immunoprecipitation suggested that CHC formed a complex with GAK and polo-like kinase 1 (PLK1). Depletion of GAK using siRNA induced metaphase arrest and aberrant localization of CHC-pT606, which abolished Kiz-pT379 (as a phosphorylation target of PLK1) signals on chromatin at metaphase. Taken together, we propose that the GAK_CHC-pT606_PLK1_Kiz-pT379 axis plays a role in proliferation of cancer cells.