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A kinetoplastid-specific kinesin is required for cytokinesis and for maintenance of cell morphology in Trypanosoma brucei

机译:布鲁氏锥虫的胞质分裂和维持细胞形态需要动素体特异性驱动蛋白

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摘要

Kinesins are motor-based transport proteins that play diverse roles in various cellular processes. The trypanosome genome lacks the homologues of many conserved mitotic kinesins, but encodes a number of trypanosome-specific kinesins with unknown function. Here, we report the biochemical and functional characterization of TbKIN-C, a trypanosome-specific kinesin, which was initially identified through an RNAi screen for cytokinesis genes in T.brucei. TbKIN-C possesses in vitro ATPase activity and associates with cytoskeletal tubulin microtubules in vivo. It is distributed throughout the cytoskeleton with a focal enrichment at the posterior end of the cell during early cell cycle stages. RNAi of TbKIN-C resulted in distorted cell shape with an elongated posterior filled with tyrosinated tubulin microtubules. Silencing of TbKIN-C impaired the segregation of organelles and cytoskeletal structures and led to detachment of the new flagellum and a small portion of the cytoplasm. We also show that RNAi of TbKIN-C compromised cytokinesis and abolished the trans-localization of TbCPC1, a subunit of the chromosomal passenger complex, from the central spindle to the initiation site of cytokinesis. Our results suggest an essential role of TbKIN-C in maintaining cell morphology, likely through regulating microtubule dynamics at the posterior end of the cell.
机译:驱动蛋白是基于马达的转运蛋白,在各种细胞过程中起着不同的作用。锥虫基因组缺乏许多保守的有丝分裂驱动蛋白的同源物,但是编码许多功能未知的锥虫特异性驱动蛋白。在这里,我们报告TbKIN-C,一种锥虫特异的驱动蛋白的生化和功能表征,最初是通过RNAi筛查布鲁氏菌中胞质分裂基因的。 TbKIN-C具有体外ATPase活性,并在体内与细胞骨架微管蛋白微管缔合。在早期细胞周期阶段,它分布在整个细胞骨架中,并在细胞的后端富集。 TbKIN-C的RNAi导致扭曲的细胞形状,后部充满酪氨酸微管蛋白微管。 TbKIN-C沉默会削弱细胞器和细胞骨架结构的分离,并导致新的鞭毛和一小部分细胞质的分离。我们还显示,TbKIN-C的RNAi破坏了胞质分裂,并取消了TbCPC1(染色体乘客复合物的一个亚基)从中心纺锤到胞质分裂起始位点的反式定位。我们的结果表明,TbKIN-C在维持细胞形态中起着至关重要的作用,可能是通过调节细胞后端的微管动力学来实现的。

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