Fission yeast ch-TOG/XMAP215 homologue Alp14 connects mitotic spindles with the kinetochore and is a component of the Mad2-dependent spindle checkpoint
Fission yeast ch-TOG/XMAP215 homologue Alp14 connects mitotic spindles with the kinetochore and is a component of the Mad2-dependent spindle checkpoint
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DOI：
10.1093/emboj/20.13.3389
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发表时间：
2001-07-02
期刊：
影响因子：
11.4
通讯作者：
Toda, T
中科院分区：
文献类型：
--
作者：
Garcia, MA;Vardy, L;Toda, T
TOG/XMAP 215相关蛋白在微管动力学的正端发挥作用。裂变酵母Alp 14是一种新发现的TOG/XMAP 215家族蛋白，与第二个同源物Dis 1一起对染色体分离至关重要。我们发现，alp 14突变体未能向正常的双极纺锤体形成的进展。有趣的是，Alp 14本身是Mad 2依赖性纺锤体检查点级联的一个组成部分，因为在添加微管去稳定化药物后，alp 14突变体无法维持高H1激酶活性，这导致securin破坏和过早的染色体分离。Alp 14-绿色荧光蛋白的实时成像显示，在有丝分裂期间，Alp 14与动粒的外围区域以及纺锤体极相关。这得到了ChIP（染色质免疫沉淀）和与动粒标记Mis 6重叠定位的支持。这些结果表明TOG/XMAP 215家族可能在着丝粒和极端到染色体微管之间起着桥梁作用。
The TOG/XMAP215-related proteins play a role in microtubule dynamics at its plus end. Fission yeast Alp14, a newly identified TOG/XMAP215 family protein, is essential for proper chromosome segregation in concert with a second homologue Dis1. We show that the alp14 mutant fails to progress towards normal bipolar spindle formation. Intriguingly, Alp14 itself is a component of the Mad2-dependent spindle checkpoint cascade, as upon addition of microtubule-destabilizing drugs the alp14 mutant is incapable of maintaining high H1 kinase activity, which results in securin destruction and premature chromosome separation. Live imaging of Alp14-green fluorescent protein shows that during mitosis, Alp14 is associated with the peripheral region of the kinetochores as well as with the spindle poles. This is supported by ChIP (chromatin immunoprecipitation) and overlapping localization with the kinetochore marker Mis6. An intact spindle is required for Alp14 localization to the kinetochore periphery, but not to the poles, These results indicate that the TOG/XMAP215 family may play a central role as a bridge between the kinetochores and the plus end of pole to chromosome microtubules.