Maiato Helder, Fairley Elizabeth A L, Rieder Conly L, Swedlow Jason R, Sunkel Claudio E, Earnshaw William C
Chromosome Structure Group, Wellcome Trust Centre for Cell Biology, Institute of Cell and Molecular Biology, University of Edinburgh, Swann Building, King's Buildings, Mayfield Road, EH9 3JR, Scotland, Edinburgh, United Kingdom.
Cell. 2003 Jun 27;113(7):891-904. doi: 10.1016/s0092-8674(03)00465-3.
One of the most intriguing aspects of mitosis is the ability of kinetochores to hold onto plus ends of microtubules that are actively gaining or losing tubulin subunits. Here, we show that CLASP1, a microtubule-associated protein, localizes preferentially near the plus ends of growing spindle microtubules and is also a component of a kinetochore region that we term the outer corona. A truncated form of CLASP1 lacking the kinetochore binding domain behaves as a dominant negative, leading to the formation of radial arrays of microtubule bundles that are highly resistant to depolymerization. Microinjection of CLASP1-specific antibodies suppresses microtubule dynamics at kinetochores and throughout the spindle, resulting in the formation of monopolar asters with chromosomes buried in the interior. Incubation with microtubule-stabilizing drugs rescues the kinetochore association with microtubule plus ends at the periphery of the asters. Our data suggest that CLASP1 is required at kinetochores for attached microtubules to exhibit normal dynamic behavior.
有丝分裂最引人入胜的一个方面是动粒能够附着在正在积极添加或丢失微管蛋白亚基的微管正端。在此,我们表明,微管相关蛋白CLASP1优先定位于正在生长的纺锤体微管的正端附近,并且也是我们称为外冠的动粒区域的一个组成部分。缺乏动粒结合结构域的CLASP1截短形式表现为显性负性,导致形成对解聚具有高度抗性的微管束径向阵列。显微注射CLASP1特异性抗体可抑制动粒处和整个纺锤体中的微管动力学,导致形成单极星状体,染色体埋在内部。用微管稳定药物孵育可挽救星状体周边动粒与微管正端的结合。我们的数据表明,动粒处需要CLASP1才能使附着的微管表现出正常的动态行为。