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果蝇 Klp67A 结合前期动粒,随后调节着丝粒聚集和纺锤体长度。

Drosophila Klp67A binds prophase kinetochores to subsequently regulate congression and spindle length.

机构信息

University of Cambridge, Department of Genetics, Cambridge, CB2 3EH, UK.

出版信息

J Cell Sci. 2010 Mar 1;123(Pt 5):767-76. doi: 10.1242/jcs.055905. Epub 2010 Feb 9.

Abstract

The kinesin-8 proteins are a family of microtubule-depolymerising motor molecules, which, despite their highly conserved roles in chromosome alignment and spindle dynamics, remain poorly characterised. Here, we report that the Drosophila kinesin-8 protein, Klp67A, exists in two spatially and functionally separable metaphase pools: at kinetochores and along the spindle. Fixed and live-cell analyses of different Klp67A recombinant variants indicate that this kinesin-8 first collects at kinetochores during prophase and, by metaphase, localises to the kinetochore outerplate. Although the catalytic motor activity of Klp67A is required for efficient kinetochore recruitment at all times, microtubules are entirely dispensable for this process. The tail of Klp67A does not play a role in kinetochore accumulation, but is both necessary and sufficient for spindle association. Using functional assays, we reveal that chromosome position and spindle length are determined by the microtubule-depolymerising motor activity of Klp67A exclusively when located at kinetochores, but not along the spindle. These data reveal that, unlike other metazoan kinesin-8 proteins, Klp67A binds the nascent prophase and mature metaphase kinetochore. From this location, Klp67A uses its motor activity to ensure chromosome alignment and proper spindle length.

摘要

驱动蛋白-8 蛋白是一类微管解聚的分子马达,尽管它们在染色体排列和纺锤体动力学中具有高度保守的作用,但仍未得到充分描述。在这里,我们报告说,果蝇的驱动蛋白-8 蛋白 Klp67A 存在于两个空间和功能上可分离的中期池:在动粒和纺锤体上。对不同 Klp67A 重组变体的固定和活细胞分析表明,这种驱动蛋白-8 首先在前期聚集在动粒上,到中期时,定位于动粒外板。尽管 Klp67A 的催化马达活性在任何时候都需要有效地招募动粒,但微管对这个过程完全是可有可无的。Klp67A 的尾部在动粒的积累过程中不起作用,但对纺锤体的结合是必需的和充分的。通过功能测定,我们揭示了染色体位置和纺锤体长度是由 Klp67A 的微管解聚马达活性决定的,这种活性仅在位于动粒上时起作用,而不在纺锤体上起作用。这些数据表明,与其他后生动物的驱动蛋白-8 蛋白不同,Klp67A 结合了早期的前期和成熟的中期动粒。从这个位置,Klp67A 利用其马达活性来确保染色体的排列和适当的纺锤体长度。

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