CDK-dependent phosphorylation and nuclear exclusion coordinately control kinetochore assembly state.
CDK-dependent phosphorylation and nuclear exclusion coordinately control kinetochore assembly state.
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DOI：
10.1083/jcb.201301006
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发表时间：
2013-04-01
期刊：
影响因子：
--
通讯作者：
Cheeseman IM
中科院分区：
文献类型：
--
作者：
Gascoigne KE;Cheeseman IM
准确的细胞分裂需要及时地分解着丝粒，这是由靶基因CDK磷酸化（包括CENP-T）和Ndc80复合体的核排斥促进的。准确的染色体分离需要多蛋白着丝点复合体的组装。先前的工作已经确定了人类细胞中100多种不同的着丝点成分。然而，关于它们在有丝分裂进入时组装和在有丝分裂退出时拆卸的调控过程，我们知之甚少。在本文中，我们使用了一种基于活细胞成像的方法来量化着丝粒拆卸动力学，并系统地分析了潜在的调节机制在控制着丝粒组装状态中的作用。我们发现着丝粒的组装和拆卸主要是由细胞周期蛋白依赖性激酶（CDK）下游的有丝分裂磷酸化驱动的。此外，我们证明细胞核排除Ndc80复合体有助于限制着丝点形成有丝分裂。结合cdk依赖性CENP-T的组成性磷酸化和Ndc80复合体的强制核定位，部分阻止了有丝分裂退出时着丝点的解体，并导致随后分裂时染色体分离缺陷。总之，我们发现外部着丝点组装的协调时间调节对于精确的细胞分裂是必不可少的。
Accurate cell division requires timely kinetochore disassembly, which is promoted by CDK phosphorylation of target genes, including CENP-T, and nuclear exclusion of the Ndc80 complex. Accurate chromosome segregation requires assembly of the multiprotein kinetochore complex. Prior work has identified more than 100 different kinetochore components in human cells. However, little is known about the regulatory processes that specify their assembly upon mitotic entry and disassembly at mitotic exit. In this paper, we used a live-cell imaging–based assay to quantify kinetochore disassembly kinetics and systematically analyze the role of potential regulatory mechanisms in controlling kinetochore assembly state. We find that kinetochore assembly and disassembly was driven primarily by mitotic phosphorylation downstream of cyclin-dependent kinase (CDK). In addition, we demonstrate that nuclear exclusion of the Ndc80 complex helped restrict kinetochore formation to mitosis. Combining constitutive CDK-dependent phosphorylation of CENP-T and forced nuclear localization of the Ndc80 complex partially prevented kinetochore disassembly at mitotic exit and led to chromosome segregation defects in subsequent divisions. In total, we find that the coordinated temporal regulation of outer kinetochore assembly is essential for accurate cell division.
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DOI：
10.1083/jcb.201001013
发表时间：
2010-06-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Carroll CW;Milks KJ;Straight AF
通讯作者：
Straight AF
影响因子：
11.8
作者：
Schmidt, Jens C.;Arthanari, Haribabu;Boeszoermenyi, Andras;Dashkevich, Natalia M.;Wilson-Kubalek, Elizabeth M.;Monnier, Nilah;Markus, Michelle;Oberer, Monika;Milligan, Ron A.;Bathe, Mark;Wagner, Gerhard;Grishchuk, Ekaterina L.;Cheeseman, Iain M.
通讯作者：
Cheeseman, Iain M.
影响因子：
8.5
作者：
Li, L.;Zhou, Y.;Fu, Y. -C.
通讯作者：
Fu, Y. -C.
影响因子：
64.5
作者：
Hori, Tetsuya;Amano, Miho;Fukagawa, Tatsuo
通讯作者：
Fukagawa, Tatsuo
影响因子：
4
作者：
Hellwig, Daniela;Emmerth, Stephan;Diekmann, Stephan
通讯作者：
Diekmann, Stephan