The human Nup107-160 nuclear pore subcomplex contributes to proper kinetochore functions
The human Nup107-160 nuclear pore subcomplex contributes to proper kinetochore functions
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DOI：
10.1038/sj.emboj.7601642
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发表时间：
2007-04-04
期刊：
影响因子：
11.4
通讯作者：
Doye, Valerie
中科院分区：
文献类型：
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作者：
Zuccolo, Michela;Alves, Annabelle;Doye, Valerie
我们之前证明了人类Nup107-160核孔亚复合物的一部分在有丝分裂开始时被招募到着丝点。然而，其着丝点定位的分子决定因素及其定位的功能意义尚未得到研究。在这里，我们发现Nup107-160复合物与CENP-F相互作用，但CENP-F仅适度地促进其靶向着丝点。此外，我们发现Nup107-160复合体对着丝点的招募主要依赖于Ndc80复合体。我们进一步证明了Nup107-160复合体从着丝点上的有效耗散，无论是通过结合靶向Seh1以外的几个亚基的sirna，还是通过单独耗散Seh1，都会诱导有丝分裂延迟。对seh1缺失细胞的进一步分析显示染色体连接受损，着丝点张力降低，着丝点-微管附着缺陷。最后，我们证明了Nup107-160复合体在着丝点上的存在是将Crm1和RanGAP1-RanBP2招募到这些结构中所必需的。总之，我们的数据因此提供了人类Nup107-160复合体在着丝点上功能的第一个分子线索。
We previously demonstrated that a fraction of the human Nup107-160 nuclear pore subcomplex is recruited to kinetochores at the onset of mitosis. However, the molecular determinants for its kinetochore targeting and the functional significance of this localization were not investigated. Here, we show that the Nup107-160 complex interacts with CENP-F, but that CENP-F only moderately contributes to its targeting to kinetochores. In addition, we show that the recruitment of the Nup107-160 complex to kinetochores mainly depends on the Ndc80 complex. We further demonstrate that efficient depletion of the Nup107-160 complex from kinetochores, achieved either by combining siRNAs targeting several of its subunits excluding Seh1, or by depleting Seh1 alone, induces a mitotic delay. Further analysis of Seh1-depleted cells revealed impaired chromosome congression, reduced kinetochore tension and kinetochore-microtubule attachment defects. Finally, we show that the presence of the Nup107-160 complex at kinetochores is required for the recruitment of Crm1 and RanGAP1-RanBP2 to these structures. Together, our data thus provide the first molecular clues underlying the function of the human Nup107-160 complex at kinetochores.