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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Examining the dynamics of chromosomal passenger complex (CPC)-dependent phosphorylation during cell division
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Examining the dynamics of chromosomal passenger complex (CPC)-dependent phosphorylation during cell division

机译：检查细胞分裂过程中依赖于染色体的客运复合体（CPC）磷酸化的动力学

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摘要

The dynamic cellular reorganization needed for successful mitosis requires regulatory cues that vary across microns. The chromo somal passenger complex (CPC) is a conserved regulator involved in key mitotic events such as chromosome-microtubule attachment and spindle midzone formation. Recently, spatial phosphoryla tion gradients have been reported for CPC substrates, raising the possibility that CPC-dependent signaling establishes order on the micron-length scale in dividing cells. However, this hypothesis has not been tested, largely because of incomplete characteriza tion of the CPC-dependent phosphorylation dynamics. Without these data it is difficult to evaluate perturbations of CPC signaling and select one that alters the spatial organization of substrate phosphorylation at a particular stage of mitosis, without changing overall phosphorylation levels. Here we examine the spatiotem poral dynamics of CPC-dependent phosphorylation along microtu bules throughout mitosis using a Forster resonance energy transfer-based sensor. We find that a CPC substrate phosphoryla tion gradient, with highest phosphorylation levels between the two spindle poles, emerges when a cell enters mitosis. Interest ingly, this gradient becomes undetectable at metaphase, but can be revealed by partially suppressing CPC activity, suggesting that high substrate phosphorylation levels can mask persistent CPC-de pendent spatial patterning. After anaphase onset, the gradient emerges and persists until cell cleavage. Selective mislocalization of the CPC during anaphase suppresses gradient formation, but overall substrate phosphorylation levels remain unchanged. Under these conditions, the spindle midzone fails to organize and func tion properly. Our findings suggest a model in which the CPC establishes phosphorylation gradients to coordinate the spatio temporal dynamics needed for error-free cell division.
机译：成功的有丝分裂所需的动态细胞重组需要调控提示，其跨微米变化。染色体乘客复合体（CPC）是一种保守的调节子，参与关键的有丝分裂事件，例如染色体-微管附着和纺锤体中间区形成。最近，已经报道了CPC底物的空间磷酸化梯度，这增加了CPC依赖性信号传导在分裂细胞中以微米长度尺度建立顺序的可能性。但是，该假设尚未得到验证，主要是因为依赖于CPC的磷酸化动力学特性不完整。没有这些数据，很难评估CPC信号的扰动并选择一个在有丝分裂的特定阶段改变底物磷酸化空间结构的方法，而又不改变总体磷酸化水平。在这里，我们使用基于Forster共振能量转移的传感器，研究了整个有丝分裂中沿着小管的CPC依赖性磷酸化的时空动态。我们发现，当细胞进入有丝分裂时，在两个纺锤极之间具有最高磷酸化水平的CPC底物磷酸化梯度出现了。有趣的是，该梯度在中期变得不可检测，但可以通过部分抑制CPC活性来揭示，这表明高底物磷酸化水平可以掩盖持续的CPC依赖的空间图案。后期开始后，梯度出现并持续直至细胞分裂。后期CPC的选择性错位抑制了梯度的形成，但总的底物磷酸化水平保持不变。在这种情况下，主轴中间区域无法正确组织和运行。我们的发现提出了一个模型，其中CPC建立了磷酸化梯度，以协调无错误细胞分裂所需的时空动态。

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