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

机译:外线粒体蛋白KNL-1在线虫中包含一个定义的寡聚域

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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 Caenorhabditis remanei oligomerizes into a decameric assembly that appears roughly circular when visualized by electron microscopy. On the basis of 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,但高阶动线粒的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以寡聚体的形式存在,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。在序列和突变分析的基础上,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的定位或导致胚胎存活力的丧失。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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