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Molecular architecture of a kinetochore–microtubule attachment site

Nature Cell Biology volume 8, pages 581–585 (2006)Cite this article

Abstract

Kinetochore attachment to spindle microtubule plus-ends is necessary for accurate chromosome segregation during cell division in all eukaryotes. The centromeric DNA of each chromosome is linked to microtubule plus-ends by eight structural-protein complexes1,2,3,4,5,6,7,8,9. Knowing the copy number of each of these complexes at one kinetochore–microtubule attachment site is necessary to understand the molecular architecture of the complex, and to elucidate the mechanisms underlying kinetochore function. We have counted, with molecular accuracy, the number of structural protein complexes in a single kinetochore–microtubule attachment using quantitative fluorescence microscopy of GFP-tagged kinetochore proteins in the budding yeast Saccharomyces cerevisiae. We find that relative to the two Cse4p molecules in the centromeric histone1, the copy number ranges from one or two for inner kinetochore proteins such as Mif2p2, to 16 for the DAM–DASH complex8,9 at the kinetochore–microtubule interface. These counts allow us to visualize the overall arrangement of a kinetochore–microtubule attachment. As most of the budding yeast kinetochore proteins have homologues in higher eukaryotes, including humans, this molecular arrangement is likely to be replicated in more complex kinetochores that have multiple microtubule attachments.

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Figure 1: Localization and turnover of kinetochore proteins in metaphase and in anaphase–telophase.
Figure 2: Characterization of the intensity distribution of a kinetochore cluster.
Figure 3: Linearity and sensitivity of the measurement technique.
Figure 4: A schematic representation of the possible architecture of a kinetochore–microtubule attachment site in metaphase.

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Acknowledgements

We thank A. Hunt, D. Odde, S. Inoué, and members of the Salmon and Bloom laboratory for helpful comments on the manuscript. This work was supported by National Institutes of Health (NIH) grants to K.S.B. (GM32238), and to E.D.S. (GM24364 and GM60678).

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  1. Department of Biology, University of North Carolina, 607 Fordham hall, CB# 3280, Chapel Hill, 27599, NC, USA

    Ajit P. Joglekar, David C. Bouck, Jeffrey N. Molk, Kerry S. Bloom & Edward D. Salmon

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Correspondence to Edward D. Salmon.

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Supplementary Information

Supplementary Notes, Tables S1, S2, S3 and S4, Figures S1, S2, S3, S4, S5 and S6 (PDF 429 kb)

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Joglekar, A., Bouck, D., Molk, J. et al. Molecular architecture of a kinetochore–microtubule attachment site. Nat Cell Biol 8, 581–585 (2006). https://doi.org/10.1038/ncb1414

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