The structure of purified kinetochores reveals multiple microtubule-attachment sites


Chromosomes must be accurately partitioned to daughter cells to prevent aneuploidy, a hallmark of many tumors and birth defects. Kinetochores are the macromolecular machines that segregate chromosomes by maintaining load-bearing attachments to the dynamic tips of microtubules. Here, we present the structure of isolated budding-yeast kinetochore particles, as visualized by EM and electron tomography of negatively stained preparations. The kinetochore appears as an ~126-nm particle containing a large central hub surrounded by multiple outer globular domains. In the presence of microtubules, some particles also have a ring that encircles the microtubule. Our data, showing that kinetochores bind to microtubules via multivalent attachments, lay the foundation to uncover the key mechanical and regulatory mechanisms by which kinetochores control chromosome segregation and cell division.

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Figure 1: Kinetochore particles contain a central hub surrounded by a number of globular domains.
Figure 2: Kinetochore particles bound to taxol-stabilized microtubules.
Figure 3: Three-dimensional structures of two types of kinetochore particles bound to a microtubule.
Figure 4: Schematic of the proposed model of kinetochore architecture.


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We are grateful to members of the Biggins and Gonen laboratories for valuable discussions and for comments on the manuscript. We are also grateful to C. Asbury, A. Powers, B. Stoddard and J. Al-Bassam for discussion and comments on the manuscript. This work was supported by US National Institutes of Health grants (GM078079 and GM064386 to S.B.), a US National Cancer Institute Cancer Center Support grant (CA015704 to S.B.) and the Howard Hughes Medical Institute (T.G.).

Author information

All authors contributed to designing the research. B.A., N.D. and S.B. performed the kinetochore purifications. S.G., M.G.I. and D.S. collected the EM data and did the EM analysis. S.B. and S.G. performed the microtubule-binding experiments. T.G. and S.B. analyzed the data and wrote the manuscript.

Correspondence to Sue Biggins or Tamir Gonen.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1 and 2, Supplementary Tables 1–4 (PDF 1766 kb)

Supplementary Movie 1

Three-dimensional tomographic reconstructions of two representative kinetochore complexes bound to microtubules. (MPG 7994 kb)

Supplementary Movie 2

Three-dimensional tomographic reconstructions of two representative kinetochore complexes bound to microtubules. (MPG 16415 kb)

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Gonen, S., Akiyoshi, B., Iadanza, M. et al. The structure of purified kinetochores reveals multiple microtubule-attachment sites. Nat Struct Mol Biol 19, 925–929 (2012).

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