Garcia Miguel Angel, Koonrugsa Nirada, Toda Takashi
Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
EMBO J. 2002 Nov 15;21(22):6015-24. doi: 10.1093/emboj/cdf611.
Fission yeast Klp5 and Klp6 belong to the microtubule-destabilizing Kin I family. In klp5 mutants, spindle checkpoint proteins Mad2 and Bub1 are recruited to mitotic kinetochores for a prolonged duration, indicating that these kinetochores are unattached. Further analysis shows that there are kinetochores to which only Bub1, but not Mad2, localizes. These kinetochores are likely to have been captured, yet lack tension. Thus Klp5 and Klp6 play a role in a spindle- kinetochore interaction at dual steps, capture and generation of tension. The TOG/XMAP215 family, Alp14 and Dis1 are known to stabilize microtubules and be required for the bivalent attachment of the kinetochore to the spindle. Despite apparent opposing activities towards microtubule stability, Klp5/Klp6 and Alp14/Dis1 share an essential function, as either dis1klp or alp14klp mutants are synthetically lethal, like alp14dis1. Defective phenotypes are similar to each other, characteristic of attachment defects and chromosome mis-segregation. Furthermore Alp14 is of significance for kinetochore localization of Klp5. We propose that Klp5/Klp6 and Alp14/Dis1 play a collaborative role in bipolar spindle formation during prometaphase through producing spindle dynamism.
裂殖酵母Klp5和Klp6属于使微管不稳定的Kin I家族。在klp5突变体中,纺锤体检查点蛋白Mad2和Bub1会在有丝分裂动粒上持续募集较长时间,这表明这些动粒未附着。进一步分析表明,存在仅定位有Bub1而没有Mad2的动粒。这些动粒可能已被捕获,但缺乏张力。因此,Klp5和Klp6在纺锤体 - 动粒相互作用的两个步骤,即捕获和张力产生中发挥作用。已知TOG/XMAP215家族的Alp14和Dis1可稳定微管,并且是动粒与纺锤体二价附着所必需的。尽管Klp5/Klp6和Alp14/Dis1对微管稳定性表现出明显相反的作用,但它们具有一项基本功能,因为dis1klp或alp14klp突变体与alp14dis1一样都是合成致死的。缺陷表型彼此相似,具有附着缺陷和染色体错分离的特征。此外,Alp14对Klp5在动粒上的定位具有重要意义。我们提出,Klp5/Klp6和Alp14/Dis1在前期通过产生纺锤体动态变化在双极纺锤体形成过程中发挥协同作用。