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Analysis of the distribution of the kinetochore protein Ndc10p in Saccharomyces cerevisiae using 3-D modeling of mitotic spindles

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Abstract

Ndc10p is one of the DNA-binding constituents of the kinetochore in Saccharomyces cerevisiae but light microscopy analysis suggests that Ndc10p is not limited to kinetochore regions. We examined the localization of Ndc10p using immunoelectron microscopy and showed that Ndc10p is associated with spindle microtubules from S-phase through anaphase. By serial section reconstruction of mitotic spindles combined with immunogold detection, we showed that Ndc10p interacts with microtubules laterally as well as terminally. About 50% of the gold label in serial section reconstructions of short mitotic spindles was associated with the walls of spindle microtubules. Interaction of kinetochore components with microtubule walls was also shown for kinetochore protein Ndc80p. Our data suggest that at least a subset of kinetochore-associated protein is dispersed throughout the mitotic spindle.

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Acknowledgements

We thank Dr. J. Kilmartin for providing us with the antibody against Ndc80p. We are grateful to Dr. K. McDonald for sharing expertise in high pressure freezing and freeze substitution. We thank Dr. F. Gail (Marine Laboratory, University of Paris 6, Paris, France) for use of the high pressure freezer and J.-P. Lechaire for expert technical assistance. We especially thank Drs. A. Desai, R. Heald, K. McDonald, J.R. McIntosh, J. Paluh, and T. U. Tanaka for their critical comments on the manuscript. Part of this work was supported by the German Research Foundation (DFG grant Mü 1423/1-1 to T. M.-R.) and by the National Institute of Health (grant RR-0592 to J.R. McIntosh). The experiments described in this paper comply with the current laws of the countries in which the experiments were performed.

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Authors and Affiliations

  1. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany

    Thomas Müller-Reichert, Anthony J. Ashford & Anthony A. Hyman

  2. European Molecular Biology Laboratory, Meyerhofstrasse 1, Postfach 10.2209, 69012 Heidelberg, Germany

    Ingrid Sassoon

  3. Boulder Laboratory for 3-D Fine Structure, Department of Molecular Cellular and Developmental Biology, University of Colorado, Boulder, CO 80309-0347, USA

    Eileen O'Toole

  4. Institut Curie UMR144 CNRS, 26 rue d'Ulm, 75248 Paris cedex 05, France

    Maryse Romao & Claude Antony

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  1. Thomas Müller-Reichert
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Correspondence to Thomas Müller-Reichert.

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Müller-Reichert, T., Sassoon, I., O'Toole, E. et al. Analysis of the distribution of the kinetochore protein Ndc10p in Saccharomyces cerevisiae using 3-D modeling of mitotic spindles. Chromosoma 111, 417–428 (2003). https://doi.org/10.1007/s00412-002-0220-6

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  • DOI: https://doi.org/10.1007/s00412-002-0220-6

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