Centriole and PCM cooperatively recruit CEP192 to spindle poles to promote bipolar spindle assembly.

Centriole and PCM cooperatively recruit CEP192 to spindle poles to promote bipolar spindle assembly.
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DOI:
10.1083/jcb.202006085
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发表时间:
2021-02-01
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Kitagawa D
Kitagawa D
中科院分区:
其他
文献类型:
--
作者:
Chinen T;Yamazaki K;Hashimoto K;Fujii K;Watanabe K;Takeda Y;Yamamoto S;Nozaki Y;Tsuchiya Y;Takao D;Kitagawa D

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通过在人类细胞系中系统地消耗PCM蛋白和中心粒,我们揭示了中心粒和PCM支架蛋白、中心粒周围蛋白和CDK5RAP2在CEP192募集到纺锤极促进双极纺锤体形成中的协同作用。在有丝分裂过程中,聚集在中心粒周围的心粒周围物质(PCM)膨胀并形成微管。在这里,我们展示了中心粒和PCM支架蛋白、中心粒蛋白和CDK5RAP2在有丝分裂期间将CEP192募集到纺锤极中的合作作用。系统地去除PCM蛋白表明,CEP192,而不是中心粒周蛋白和/或CDK5RAP2,对HeLa、RPE1和A549细胞的双极性纺锤体组装至关重要。在中心蛋白和CDK5RAP2的双重耗竭下,CEP192留在中心粒壁上,足以形成双极纺锤体。相反,通过去除中心粒,我们发现中心粒蛋白和CDK5RAP2在中心粒纺锤极招募CEP192,并促进具有一个中心体的有丝分裂细胞的双极性纺锤体形成。此外,对PCM组装的关键激酶PLK1的扰动有效地抑制了具有一个中心体的有丝分裂细胞的双极纺锤体形成。总的来说,这些数据表明中心粒和PCM支架蛋白协同将CEP192招募到纺锤极并促进双极纺锤体的形成。
By systematic depletion of PCM proteins and centrioles in human cell lines, we reveal the cooperative role of the centriole and PCM scaffold proteins, pericentrin and CDK5RAP2, in the recruitment of CEP192 to spindle poles to promote bipolar spindle formation. The pericentriolar material (PCM) that accumulates around the centriole expands during mitosis and nucleates microtubules. Here, we show the cooperative roles of the centriole and PCM scaffold proteins, pericentrin and CDK5RAP2, in the recruitment of CEP192 to spindle poles during mitosis. Systematic depletion of PCM proteins revealed that CEP192, but not pericentrin and/or CDK5RAP2, was crucial for bipolar spindle assembly in HeLa, RPE1, and A549 cells with centrioles. Upon double depletion of pericentrin and CDK5RAP2, CEP192 that remained at centriole walls was sufficient for bipolar spindle formation. In contrast, through centriole removal, we found that pericentrin and CDK5RAP2 recruited CEP192 at the acentriolar spindle pole and facilitated bipolar spindle formation in mitotic cells with one centrosome. Furthermore, the perturbation of PLK1, a critical kinase for PCM assembly, efficiently suppressed bipolar spindle formation in mitotic cells with one centrosome. Overall, these data suggest that the centriole and PCM scaffold proteins cooperatively recruit CEP192 to spindle poles and facilitate bipolar spindle formation.
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