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Kinetochores distinguish GTP from GDP forms of the microtubule lattice

Nature volume 388, pages 888–891 (1997)Cite this article

Abstract

During prometaphase in mitotic cell division, chromosomes attach to the walls of microtubules and subsequently move to microtubule ends, where they stay throughout mitosis1,2. This end-attachment seems to be essential for correct chromosome segregating. However, the mechanism by which kinetochores, the multiprotein complexes that link chromosomes to the microtubules of the mitotic spindle3,4, recognize and stay attached to microtubule ends is not understood. One clue comes from the hydrolysis of GTP that occurs during microtubule polymerization. Although tubulin dimers must contain GTP to polymerize, this GTP is rapidly hydrolysed following the addition of dimers to a growing polymer. This creates a microtubule consisting largely of GDP-tubulin, with a small cap of GTP-tubulin at the end5. It is possible that kinetochores distinguish the different structural states of a GTP- versus a GDP-microtubule lattice. We have examined this question in vitro using reconstituted kinetochores from the yeast Saccharomyces cerevisiae. We found that kinetochores in vitro bind preferentially to GTP- rather than GDP-microtubules, and to the plus-end preferentially over the lattice. Our results could explain how kinetochores stay at microtubule ends and thus segregate chromosomes correctly during mitosis invivo. This result demonstrates that proteins exist that can distinguish the GTP conformation of the microtubule lattice.

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Figure 1: a, Preparation of segmented microtubules.
Figure 2: a, Photograph of the capped microtubule with the bead bound to the GMPCPP tubulin region.
Figure 3: a, The bead bound to the plus-end of the GMPCPP microtubule.
Figure 4: Model for kinetochore behaviour on mitotic-spindle microtubule.

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Acknowledgements

We thank S. Schuyler for early experiments suggesting the role of lattice structure; J.Howard for help with statistics; and M. Glotzer, P. Gonczy, S. Eaton, K. Simons, R. Heald, S. Reinsch and S. Andersen for critical reading of the manuscript.

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Authors and Affiliations

  1. European Molecular Biology Laboratory,, Postfach, 10.2209, D-69012, Heidelberg, Germany

    Fedor F. Severin & Anthony A. Hyman

  2. Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Peter K. Sorger

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  2. Peter K. Sorger
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  3. Anthony A. Hyman
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Severin, F., Sorger, P. & Hyman, A. Kinetochores distinguish GTP from GDP forms of the microtubule lattice. Nature 388, 888–891 (1997). https://doi.org/10.1038/42270

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