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Cnn1 inhibits the interactions between the KMN complexes of the yeast kinetochore

Nature Cell Biology volume 14, pages 614–624 (2012)Cite this article

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A Corrigendum to this article was published on 01 March 2013

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Abstract

Kinetochores attach the replicated chromosomes to the mitotic spindle and orchestrate their transmission to the daughter cells. Kinetochore–spindle binding and chromosome segregation are mediated by the multi-copy KNL1Spc105, MIS12Mtw1 and NDC80Ndc80 complexes that form the so-called KMN network. KMN–spindle attachment is regulated by the Aurora BIpl1 and MPS1Mps1 kinases. It is unclear whether other mechanisms exist that support KMN activity during the cell cycle. Using budding yeast, we show that kinetochore protein Cnn1 localizes to the base of the Ndc80 complex and promotes a functionally competent configuration of the KMN network. Cnn1 regulates KMN activity in a spatiotemporal manner by inhibiting the interaction between its complexes. Cnn1 activity peaks in anaphase and is driven by the Cdc28, Mps1 and Ipl1 kinases.

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Figure 1: Cnn1 is a kinetochore component whose concentration at kinetochores peaks in anaphase.
Figure 2: The enrichment of Cnn1 at anaphase kinetochores is phospho-driven.
Figure 3: Cnn1 protein–protein interactions and localization to the inner kinetochore.
Figure 4: Cnn1 ensures a timely progression through S phase and promotes faithful chromosome transmission.
Figure 5: Cnn1 promotes the sister-chromatid/spindle binding and bi-orientation activity of the KMN network.
Figure 6: Cnn1 inhibits the interaction between the KMN complexes.

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  • 11 February 2013

    In the version of this Article that was originally published, the acknowledgement to M. Winey and reference 48 in the Methods section were omitted in error. These corrections have been made in the PDF and HTML versions of the Article.

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Acknowledgements

The authors thank A. Amon (MIT, USA), S. Biggins (University of Washington, USA), D. Koshland (University of California, Berkeley, USA), M. Singleton (Cancer Research UK), F. Solomon (MIT, USA), P. K. Sorger (Harvard Medical School, USA) and M. Winey (University of Colorado, USA) for strains or reagents. We also thank M. Foiani, S. Ghandi, D. Lamont, J. Lechner, C. Lucca, J. Ortiz, S. ten Have and R. Visintin for technical support or discussions. We thank A. Schleiffer and S. Westermann for communicating results before publication. P.D.W. acknowledges financial support from the Italian Association for Cancer Research (grant 8840). T.R.H. recognizes support from the N.I.H. (grant GM087461) and the American Cancer Society (grant IRG 58-006-50). T.U.T. acknowledges a Cancer Research U.K. senior fellowship and Wellcome Trust program grant. L.J.B. acknowledges a doctoral fellowship from the European School of Molecular Medicine.

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  1. Lucy J. Bock and Cinzia Pagliuca: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Experimental Oncology, European Institute of Oncology, Via Adamello 16, 20139 Milan, Italy

    Lucy J. Bock, Cinzia Pagliuca, Claudio Alfieri, Cristina Golfieri, Marianna Dal Maschio & Peter De Wulf

  2. Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, UK

    Norihiko Kobayashi, Yusuke Oku & Tomoyuki U. Tanaka

  3. Department of Medicinal Chemistry and Molecular Pharmacology, Center for Cancer Research, Purdue University, West Lafayette, Indiana 47907-2091, USA

    Ryan A. Grove, Kriti Shrestha & Tony R. Hazbun

  4. FIRC Institute of Molecular Oncology Foundation, 20139 Milan, Italy

    Amanda Oldani

  5. Instituto de Biología Funcional y Genómica, CSIC, Universidad de Salamanca, 37008 Salamanca, Spain

    Rodrigo Bermejo

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Contributions

L.J.B. and C.P. performed all the genetic, molecular biological, biochemical and cell biological experiments. N.K. performed the time-lapse CEN-reactivation and Cnn1–3GFP imaging experiments, R.A.G. performed the Y2H screens, Y.O. performed the in vitro kinase experiments, K.S. performed the in vitro Cnn1–KMN binding experiments, C.A. helped with the genetic interaction screens and the ChIP–chip experiment, C.G. and M.D.M. performed the cell-cycle experiments, PhosTag western hybridization and Cnn1–Cnn1 co-immunoprecipitation experiments. A.O. performed the FRAP experiments and helped with the quantification of Cnn1–3GFP and Ndc80–3GFP at kinetochores, R.B. helped with the ChIP–chip experiment and converted the microarray data into graphic format, T.R.H. supervised the Y2H screens and the in vitro Cnn1–KMN binding experiments. T.U.T. supervised the live-cell imaging experiments. P.D.W. supervised the project, helped with yeast imaging and wrote the paper.

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Correspondence to Peter De Wulf.

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Bock, L., Pagliuca, C., Kobayashi, N. et al. Cnn1 inhibits the interactions between the KMN complexes of the yeast kinetochore. Nat Cell Biol 14, 614–624 (2012). https://doi.org/10.1038/ncb2495

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