Kinetochore stretching inactivates the spindle assembly checkpoint.

Kinetochore stretching inactivates the spindle assembly checkpoint.
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DOI:
10.1083/jcb.200811028
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发表时间:
2009-02-09
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Hirota T
Hirota T
中科院分区:
其他
文献类型:
--
作者:
Uchida KS;Takagaki K;Kumada K;Hirayama Y;Noda T;Hirota T

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纺锤体组装检查点(SAC)监测微管与动粒的附着,并在微管结合不完全时抑制后期。SAC也可能对张力做出反应;然而,细胞如何感知张力以及它的检测是否对满足SAC很重要仍然存在争议。我们产生了一个HeLa细胞系,其中着丝粒蛋白A和Mis 12的两个组成部分,分别用绿色和红色荧光团标记。这些细胞的活细胞成像揭示了双取向后动粒延伸和反冲的重复循环。在动粒伸展被抑制的条件下,细胞不能沉默SAC,并在延迟后进入后期,不管着丝粒伸展。监测细胞周期蛋白B水平作为后期促进复合物/细胞周期体活性的读数,我们发现,抑制动粒拉伸延迟和减速细胞周期蛋白B降解。这些观察结果表明,SAC监测着丝粒,而不是着丝粒的拉伸和着丝粒拉伸促进沉默的SAC信号。
The spindle assembly checkpoint (SAC) monitors the attachment of microtubules to the kinetochore and inhibits anaphase when microtubule binding is incomplete. The SAC might also respond to tension; however, how cells can sense tension and whether its detection is important to satisfy the SAC remain controversial. We generated a HeLa cell line in which two components of the kinetochore, centromere protein A and Mis12, are labeled with green and red fluorophores, respectively. Live cell imaging of these cells reveals repetitive cycles of kinetochore extension and recoiling after biorientation. Under conditions in which kinetochore stretching is suppressed, cells fail to silence the SAC and enter anaphase after a delay, regardless of centromere stretching. Monitoring cyclin B levels as a readout for anaphase-promoting complex/cyclosome activity, we find that suppression of kinetochore stretching delays and decelerates cyclin B degradation. These observations suggest that the SAC monitors stretching of kinetochores rather than centromeres and that kinetochore stretching promotes silencing of the SAC signal.
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