Kinesin-14s are conserved molecular motors required for high-fidelity chromosome segregation, but their specific contributions to spindle function have not been fully defined. Here, we show that key functions of budding yeast Kinesin-14 Cik1-Kar3 are accomplished in a complex with Bim1 (yeast EB1). Genetic complementation of mitotic phenotypes identifies a novel KLTF peptide motif in the Cik1 N-terminus. We show that this motif is one element of a tripartite binding interface required to form a high-affinity Bim1–Cik1-Kar3 complex. Lack of Bim1-binding by Cik1-Kar3 delays cells in mitosis and impairs microtubule bundle organization and dynamics. Conversely, constitutive targeting of Cik1-Kar3 to microtubule plus ends induces the formation of nuclear microtubule bundles. Cells lacking the Bim1–Cik1-Kar3 complex rely on the conserved microtubule bundler Ase1/PRC1 for metaphase spindle organization, and simultaneous loss of plus-end targeted Kar3 and Ase1 is lethal. Our results reveal the contributions of an EB1–Kinesin-14 complex for spindle formation as a prerequisite for efficient kinetochore clustering and bi-orientation.
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7 December 2020
Article|
October 07 2020
The EB1–Kinesin-14 complex is required for efficient metaphase spindle assembly and kinetochore bi-orientation
Nikolay Kornakov,
Nikolay Kornakov
Department of Molecular Genetics, Faculty of Biology, Center of Medical Biotechnology, University of Duisburg-Essen, Essen, Germany
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Bastian Möllers,
Bastian Möllers
Department of Molecular Genetics, Faculty of Biology, Center of Medical Biotechnology, University of Duisburg-Essen, Essen, Germany
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Stefan Westermann
Department of Molecular Genetics, Faculty of Biology, Center of Medical Biotechnology, University of Duisburg-Essen, Essen, Germany
Correspondence to Stefan Westermann: Stefan.Westermann@uni-due.de
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Nikolay Kornakov
Department of Molecular Genetics, Faculty of Biology, Center of Medical Biotechnology, University of Duisburg-Essen, Essen, Germany
Bastian Möllers
Department of Molecular Genetics, Faculty of Biology, Center of Medical Biotechnology, University of Duisburg-Essen, Essen, Germany
Stefan Westermann
Department of Molecular Genetics, Faculty of Biology, Center of Medical Biotechnology, University of Duisburg-Essen, Essen, Germany
Correspondence to Stefan Westermann: Stefan.Westermann@uni-due.de
Received:
March 12 2020
Revision Received:
July 28 2020
Accepted:
September 10 2020
Online Issn: 1540-8140
Print Issn: 0021-9525
© 2020 Kornakov et al.
2020
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Cell Biol (2020) 219 (12): e202003072.
Article history
Received:
March 12 2020
Revision Received:
July 28 2020
Accepted:
September 10 2020
Citation
Nikolay Kornakov, Bastian Möllers, Stefan Westermann; The EB1–Kinesin-14 complex is required for efficient metaphase spindle assembly and kinetochore bi-orientation. J Cell Biol 7 December 2020; 219 (12): e202003072. doi: https://doi.org/10.1083/jcb.202003072
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