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The outer kinetochore protein KNL-1 contains a defined oligomerization domain in nematodes

机译:外部动粒蛋白KNL-1在线虫中含有确定的寡聚化结构域

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摘要

The kinetochore is a large, macromolecular assembly that is essential for connecting chromosomes to microtubules during mitosis. Despite the recent identification of multiple kinetochore components, the nature and organization of the higher order kinetochore structure remain unknown. The outer kinetochore KNL-1/Mis12 complex/Ndc80 complex (KMN) network plays a key role in generating and sensing microtubule attachments. Here, we demonstrate that Caenorhabditis elegans KNL-1 exists as an oligomer and we identify a specific domain in KNL-1 responsible for this activity. An N-terminal KNL-1 domain from both C. elegans and the related nematode C. remanei oligomerizes into a decameric assembly that appears roughly circular when visualized by electron microscopy. Based on sequence and mutational analysis, we identify a small hydrophobic region as responsible for this oligomerization activity. However, mutants that precisely disrupt KNL-1 oligomerization did not alter KNL-1 localization or result in the loss of embryonic viability based on gene replacements in C. elegans. In C. elegans, KNL-1 oligomerization may coordinate with other kinetochore activities to ensure the proper organization, function, and sensory capabilities of the kinetochore-microtubule attachment.
机译:线粒体是一个大型的大分子装配体,对于在有丝分裂期间将染色体连接到微管至关重要。尽管最近发现了多个动线粒的组成部分,但高阶动线粒结构的性质和组织仍然未知。外部动粒KNL-1 / Mis12复合物/ Ndc80复合物(KMN)网络在生成和感测微管附件中起关键作用。在这里,我们证明秀丽隐杆线虫KNL-1以寡聚物形式存在,并且我们在KNL-1中鉴定了负责此活性的特定结构域。来自秀丽隐杆线虫和相关线虫秀丽隐杆线虫的N末端KNL-1结构域寡聚成十聚体组装体,当通过电子显微镜观察时,该组装体看起来大致为圆形。基于序列和突变分析,我们确定了一个小疏水区域负责这种低聚活性。然而,基于秀丽隐杆线虫的基因置换,精确破坏KNL-1寡聚化的突变体不会改变KNL-1的定位或导致胚胎生存力的丧失。在秀丽隐杆线虫中,KNL-1寡聚可能与其他动线粒活动协调,以确保动线粒-微管附件的适当组织,功能和感觉能力。

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