High-Resolution Temporal Analysis Reveals a Functional Timeline for the Molecular Regulation of Cytokinesis
High-Resolution Temporal Analysis Reveals a Functional Timeline for the Molecular Regulation of Cytokinesis
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DOI：
10.1016/j.devcel.2014.05.009
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发表时间：
2014-07-28
影响因子：
11.8
通讯作者：
Canman, Julie C.
中科院分区：
文献类型：
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作者：
Davies, Tim;Jordan, Shawn N.;Canman, Julie C.
为了充分利用快速作用的温度敏感突变，热控制必须非常迅速。我们开发了Therminator，这是一种能够在大约17秒内改变样品温度的装置，同时可以在体内成像细胞分裂。将该技术应用于细胞分裂的六个关键调节因子，我们发现每个调节因子在秀丽隐杆线虫受精卵中都有不同的时间要求。具体来说，肌球蛋白- ii在整个细胞分裂过程中都是必需的，直到收缩环关闭。相比之下，福尔明介导的肌动蛋白成核只需要在组装和早期收缩环收缩期间进行。在环闭合后，需要中央纺锤体保持分裂，尽管它的GAP活动只需要到闭合之前。最后，染色体客运复合体仅在有丝分裂早期需要细胞质分裂，而在中期或细胞质分裂时不需要。总之，我们的研究结果为使用Therminator（一种超快速蛋白质失活的强大工具）的细胞分裂分子调节因子提供了精确的功能时间表。
To take full advantage of fast-acting temperature-sensitive mutations, thermal control must be extremely rapid. We developed the Therminator, a device capable of shifting sample temperature in similar to 17 s while simultaneously imaging cell division in vivo. Applying this technology to six key regulators of cytokinesis, we found that each has a distinct temporal requirement in the Caenorhabditis elegans zygote. Specifically, myosin-II is required throughout cytokinesis until contractile ring closure. In contrast, formin-mediated actin nucleation is only required during assembly and early contractile ring constriction. Centralspindlin is required to maintain division after ring closure, although its GAP activity is only required until just prior to closure. Finally, the chromosomal passenger complex is required for cytokinesis only early in mitosis, but not during metaphase or cytokinesis. Together, our results provide a precise functional timeline for molecular regulators of cytokinesis using the Therminator, a powerful tool for ultra-rapid protein inactivation.