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The Ndc80 kinetochore complex forms oligomeric arrays along microtubules

Nature volume 467, pages 805–810 (2010)Cite this article

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Abstract

The Ndc80 complex is a key site of regulated kinetochore–microtubule attachment (a process required for cell division), but the molecular mechanism underlying its function remains unknown. Here we present a subnanometre-resolution cryo-electron microscopy reconstruction of the human Ndc80 complex bound to microtubules, sufficient for precise docking of crystal structures of the component proteins. We find that the Ndc80 complex binds the microtubule with a tubulin monomer repeat, recognizing α- and β-tubulin at both intra- and inter-tubulin dimer interfaces in a manner that is sensitive to tubulin conformation. Furthermore, Ndc80 complexes self-associate along protofilaments through interactions mediated by the amino-terminal tail of the NDC80 protein, which is the site of phospho-regulation by Aurora B kinase. The complex’s mode of interaction with the microtubule and its oligomerization suggest a mechanism by which Aurora B could regulate the stability of load-bearing kinetochore–microtubule attachments.

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Figure 1: Structure of the Ndc80 complex–microtubule interface.
Figure 2: The NDC80 toe-print is a tubulin conformation sensor.
Figure 3: Cluster formation requires the N terminus of the NDC80 protein.
Figure 4: Proposed models of attachment maturation and biased diffusion.

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Protein Data Bank

Data deposits

The cryo-electron microscopy densitymap and coordinates of the docking model have been deposited at the EMDB and PDB under accession numbers 5223 and 3IZ0, respectively.

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Acknowledgements

We are grateful to K. H. Downing for supporting the work carried out by D.A.B., to C. Ciferri for his knowledge and advice about the Ndc80 complex and critical reading of the manuscript, and to P. Grob and S. Lipscomb for electron-microscopy and computer support, respectively. We also acknowledge D. Typke and B. Glaeser for advice on data collection, and C. Sindelar for discussion of data processing strategies. This work was funded by a grant from the National Institute of General Medical Sciences (E.N.). E.N. and N.G. are Howard Hughes Medical Institute Investigators.

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Authors and Affiliations

  1. Biophysics Graduate Group, University of California, Berkeley, 94720, California, USA

    Gregory M. Alushin & Vincent H. Ramey

  2. Department of Experimental Oncology, European Institute of Oncology, 20139 Milan, Italy

    Sebastiano Pasqualato & Andrea Musacchio

  3. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, 94720, California, USA

    David A. Ball & Eva Nogales

  4. Howard Hughes Medical Institute, Rosenstiel Basic Medical Research Center, Brandeis University, Waltham, 02453, Massachusetts, USA

    Nikolaus Grigorieff

  5. Research Unit of the Italian Institute of Technology at the IFOM-IEO Campus, 20139 Milan, Italy

    Andrea Musacchio

  6. Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, 94720, California, USA

    Eva Nogales

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Contributions

G.M.A. performed research. G.M.A. and V.H.R. developed data processing tools. G.M.A. and S.P. generated and purified Ndc80(bonsai) mutants. D.A.B. generated the tomograms displayed in Supplementary Fig. 1. N.G. adapted Frealign software for helical samples and generated the final refined reconstruction. G.M.A. and E.N. designed research. All authors analysed data and discussed the results. G.M.A., A.M. and E.N. wrote the Article.

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Correspondence to Eva Nogales.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains a Supplementary Discussion, additional references, Supplementary Figures 1-13 with legends, Supplementary Table 1 and legends for Supplementary Movies 1-2. (PDF 21402 kb)

Supplementary Movie 1

This movie files shows pseudoatomic model of the Ndc80-microtubule interface - see Supplementary Information file for full legend. (MOV 12365 kb)

Supplementary Movie 2

This movie file shows serial slices of wild-type Ndc80 bonsai bound to microtubules under sub-saturating conditions - see Supplementary Information file for full legend. (MOV 2773 kb)

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Alushin, G., Ramey, V., Pasqualato, S. et al. The Ndc80 kinetochore complex forms oligomeric arrays along microtubules. Nature 467, 805–810 (2010). https://doi.org/10.1038/nature09423

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