Department of Biology, University of Washington, Seattle, WA 98195.
Department of Statistics, University of Washington, Seattle, WA 98195.
Proc Natl Acad Sci U S A. 2017 Jul 18;114(29):E5854-E5863. doi: 10.1073/pnas.1705096114. Epub 2017 Jul 5.
Devoid of all known canonical actin-binding proteins, the prevalent parasite uses an alternative mechanism for cytokinesis. Unique aspects of this mechanism can potentially be leveraged for therapeutic development. Here, live-cell imaging methods were developed for to establish division kinetics and the core division machinery. Surprisingly, cytokinesis occurred with a median time that is ∼60 times faster than mammalian cells. In contrast to cells that use a contractile ring, actin was not concentrated in the furrow and was not directly required for furrow progression. Live-cell imaging and morpholino depletion of axonemal Paralyzed Flagella 16 indicated that flagella-based forces initiated daughter cell separation and provided a source for membrane tension. Inhibition of membrane partitioning blocked furrow progression, indicating a requirement for membrane trafficking to support furrow advancement. Rab11 was found to load onto the intracytoplasmic axonemes late in mitosis and to accumulate near the ends of nascent axonemes. These developing axonemes were positioned to coordinate trafficking into the furrow and mark the center of the cell in lieu of a midbody/phragmoplast. We show that flagella motility, Rab11, and actin coordination are necessary for proper abscission. Organisms representing three of the five eukaryotic supergroups lack myosin II of the actomyosin contractile ring. These results support an emerging view that flagella play a central role in cell division among protists that lack myosin II and additionally implicate the broad use of membrane tension as a mechanism to drive abscission.
该寄生虫缺乏所有已知的经典肌动蛋白结合蛋白,因此采用替代机制进行胞质分裂。该机制的独特方面可能被用于开发治疗方法。在这里,开发了活细胞成像方法来建立 分裂动力学和核心分裂机制。令人惊讶的是,与使用收缩环的细胞相比, 胞质分裂的中位数时间快了约 60 倍。与使用收缩环的细胞不同,肌动蛋白不会在沟中浓缩,也不是沟进展所必需的。活细胞成像和轴丝 Paralyzed Flagella 16 的形态发生体耗竭表明,基于鞭毛的力启动子细胞分离,并为膜张力提供来源。抑制膜分区会阻止沟的进展,表明需要膜运输来支持沟的推进。发现 Rab11 在有丝分裂后期加载到细胞质内轴丝上,并在新形成的轴丝末端附近积累。这些正在发育的轴丝被定位以协调进入沟的运输,并取代中体/收缩环作为细胞的中心。我们表明,鞭毛运动、Rab11 和肌动蛋白协调是正确分离所必需的。代表五个真核超组中的三个的生物体缺乏肌球蛋白 II 的肌动球蛋白收缩环。这些结果支持了一个新兴观点,即鞭毛在缺乏肌球蛋白 II 的原生动物的细胞分裂中发挥核心作用,此外还暗示了广泛使用膜张力作为驱动分离的机制。