Department of Biology, University of Washington, Seattle, WA 98195.
Department of Biochemistry, University of Washington, Seattle, WA 98195.
Mol Biol Cell. 2020 Jul 15;31(15):1611-1622. doi: 10.1091/mbc.E19-07-0406. Epub 2020 May 27.
has 198 Nek kinases whereas humans have only 11. has a complex microtubule cytoskeleton that includes eight flagella and several unique microtubule arrays that are utilized for parasite attachment and facilitation of rapid mitosis and cytokinesis. The need to regulate these structures may explain the parallel expansion of the number of Nek family kinases. Here we use live and fixed cell imaging to uncover the role of Nek8445 in regulating cell division. We demonstrate that Nek8445 localization is cell cycle regulated and this kinase has a role in regulating overall microtubule organization. Nek8445 depletion results in short flagella, aberrant ventral disk organization, loss of the funis, defective axoneme exit, and altered cell shape. The axoneme exit defect is specific to the caudal axonemes, which exit from the posterior of the cell, and this defect correlates with rounding of the cell posterior and loss of the funis. Our findings implicate a role for the funis in establishing cell shape and guiding axoneme docking. On a broader scale our results support the emerging view that Nek family kinases have a general role in regulating microtubule organization.
拥有 198 个 Nek 激酶,而人类只有 11 个。它具有复杂的微管细胞骨架,包括 8 个鞭毛和几个独特的微管阵列,用于寄生虫附着和促进快速有丝分裂和胞质分裂。需要调节这些结构可能解释了 Nek 家族激酶数量的平行扩张。在这里,我们使用活细胞和固定细胞成像来揭示 Nek8445 在调节细胞分裂中的作用。我们证明 Nek8445 的定位是细胞周期调节的,这种激酶在调节整体微管组织中起作用。Nek8445 的耗竭导致鞭毛变短、腹盘组织异常、系带丢失、轴突退出缺陷和细胞形状改变。轴突退出缺陷是尾轴突特有的,尾轴突从细胞的后部退出,并且这种缺陷与细胞后部的变圆和系带的丢失相关。我们的研究结果表明,系带在建立细胞形状和引导轴突对接中起作用。从更广泛的角度来看,我们的结果支持新兴观点,即 Nek 家族激酶在调节微管组织中具有普遍作用。