Cancer Research UK Cell Cycle Genetics Research Group, Department of Genetics, University of Cambridge, Cambridge, United Kingdom.
PLoS One. 2007 Jun 27;2(6):e572. doi: 10.1371/journal.pone.0000572.
Polo-like kinases control multiple events during cell division, including mitotic entry, centrosome organization, spindle formation, chromosome segregation and cytokinesis. Their roles during cytokinesis, however, are not well understood because the requirement of these kinases during early stages of mitosis complicates the study of their functions after anaphase onset.
METHODOLOGY/PRINCIPAL FINDINGS: We used time-lapse microscopy to analyze the dynamics of Polo::GFP in Drosophila tissue culture cells during mitosis. After anaphase onset, Polo::GFP concentrated at the spindle midzone, but also diffused along the entire length of the central spindle. Using RNA interference we demonstrate that the microtubule-associated proteins Feo and Klp3A are required for Polo recruitment to the spindle midzone, but not the kinesin Pavarotti as previously thought. Moreover, we show that Feo and Klp3A form a complex and that Polo co-localizes with both proteins during cytokinesis.
CONCLUSION/SIGNIFICANCE: Our results reveal that the Feo/Klp3A complex is necessary for Polo recruitment to the spindle midzone. A similar finding has also been recently reported in mammalian cells [1], suggesting that this basic mechanism has been conserved during evolution, albeit with some differences. Finally, since cleavage furrow formation and ingression are unaffected following feo RNAi, our data imply that Polo recruitment to the central spindle is not required for furrowing, but some other aspect of cytokinesis.
Polo 样激酶在细胞分裂过程中控制着多种事件,包括有丝分裂进入、中心体组织、纺锤体形成、染色体分离和胞质分裂。然而,它们在胞质分裂中的作用还不是很清楚,因为这些激酶在有丝分裂早期的需求使得研究它们在后期开始后的功能变得复杂。
方法/主要发现:我们使用延时显微镜分析了果蝇组织培养细胞中 Polo::GFP 在有丝分裂过程中的动力学。后期开始后,Polo::GFP 集中在纺锤体中部,但也沿着中央纺锤体的全长扩散。通过 RNA 干扰,我们证明微管相关蛋白 Feo 和 Klp3A 对于 Polo 招募到纺锤体中部是必需的,但不是以前认为的驱动蛋白 Pavarotti。此外,我们还表明 Feo 和 Klp3A 形成复合物,并且在胞质分裂过程中 Polo 与这两种蛋白共定位。
结论/意义:我们的结果表明,Feo/Klp3A 复合物对于 Polo 招募到纺锤体中部是必需的。在哺乳动物细胞中最近也有类似的发现[1],这表明尽管存在一些差异,但这种基本机制在进化过程中得到了保守。最后,由于在 feo RNAi 后,分裂沟的形成和侵入不受影响,我们的数据意味着 Polo 招募到中央纺锤体对于沟的形成不是必需的,但对于胞质分裂的其他某些方面是必需的。