Article
- The EMBO Journal (2007) 26, 1853 - 1864
- doi:10.1038/sj.emboj.7601642
Published online: 15 March 2007
Subject Categories:
The human Nup107–160 nuclear pore subcomplex contributes to proper kinetochore functions
Michela Zuccolo1,2, Annabelle Alves1,2, Vincent Galy3, Stéphanie Bolhy1,2, Etienne Formstecher4, Victor Racine1,2, Jean-Baptiste Sibarita1,2, Tatsuo Fukagawa5, Ramin Shiekhattar6, Tim Yen7 and Valérie Doye1,2
- Institut Curie, Centre de Recherche, Paris, France
- UMR144 CNRS, Paris, France
- Unité de Biologie Cellulaire du Noyau, URA 2582 CNRS - Institut Pasteur, Paris, France
- HYBRIGENICS SA, Paris, France
- Department of Molecular Genetics, National Institute of Genetics and SOKENDAI, Mishima, Shizuoka, Japan
- The Wistar Institute, Philadelphia, PA, USA
- Fox Chase Cancer Center, Philadelphia, PA, USA
Correspondence to:
Valérie Doye, Institut Curie, Centre de Recherche, UMR144 CNRS, 26 rue d'Ulm, 75248 Paris Cedex 05, France. Tel.: +33 1 42 34 64 10; Fax: + 33 1 42 34 64 21; E-mail: vdoye@curie.fr
Received 1 September 2006; Accepted 14 February 2007
Abstract
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.
Keywords:
- CENP-F,
- Crm1,
- Ndc80 complex,
- nucleoporin,
- Seh1
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