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γ-Tubulin complex-mediated anchoring of spindle microtubules to spindle-pole bodies requires Msd1 in fission yeast

Nature Cell Biology volume 9, pages 646–653 (2007)Cite this article

Abstract

The anchoring of microtubules to subcellular structures is critical for cell polarity and motility. Although the process of anchoring cytoplasmic microtubules to the centrosome has been studied in some detail1,2,3,4, it is not known how spindle microtubules are anchored to the mitotic centrosome and, particularly, whether anchoring and nucleation of mitotic spindles are functionally separate. Here, we show that a fission yeast coiled-coil protein, Msd1, is required for anchoring the minus end of spindle microtubules to the centrosome equivalent, the spindle-pole body (SPB). msd1 deletion causes spindle microtubules to abnormally extend beyond SPBs, which results in chromosome missegregation. Importantly, this protruding spindle is phenocopied by the amino-terminal deletion mutant of Alp4, a component of the γ-tubulin complex5 (γ-TuC), which lacks the potential Msd1-interacting domain. We propose that Msd1 interacts with γ-TuC, thereby specifically anchoring the minus end of microtubules to SPBs without affecting microtubule nucleation.

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Figure 1: Mitosis-specific localization of Msd1.
Figure 2: Protruding mitotic spindle microtubules in the msd1 deletion mutant.
Figure 3: The minus end of spindle microtubules is no longer tethered to the SPB in the msd1 mutant.
Figure 4: Chromosome missegregation in the msd1 mutant.
Figure 5: Interaction between Msd1 and Alp4 and the protruding spindle phenotypes in alp4 mutants.

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Acknowledgements

We thank F. Uhlmann and R. E. Carazo-Salas for critical reading of the manuscript, discussion and technical support for microscopy (R. E. Carazo-Salas). We are grateful to A. McAinsh for pointing out the homology between Msd1 and TINA. We also thank F. Chang, K. Gould, K. Gull, O. Niwa, J. R. McIntosh, K. Sawin, K. Tanaka, I. Hagan, M. Yanagida and M. Yoshida for reagents and/or information, and A. Sugimoto and M. Yamamoto for support. This work was supported by Cancer Research UK (T.T.).

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Author notes

  1. Mika Toya

    Present address: Current address: Laboratory for Developmental Genetics, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan.,

  2. Masamitsu Sato

    Present address: Current address: Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan.,

  3. Kazuhide Asakawa

    Present address: Current address: Division of Molecular and Developmental Biology, Department of Developmental Genetics, National Institute of Genetics, Yata 1111, Mishima, Shizuoka 411-8540, Japan.,

Authors and Affiliations

  1. Laboratory of Cell Regulation, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Mika Toya, Masamitsu Sato, Kazuhide Asakawa & Takashi Toda

  2. Cell Biology and Biophysics Program, European Molecular Biology Laboratory, Meyerhofstrasse 1, Heidelberg, D-69117, Germany

    Uta Haselmann, Damian Brunner & Claude Antony

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Contributions

M.T. performed all experiments and M.S. provided experimental materials and made suggestions for experimental design. K.A. contributed to initial identification of Msd1. Electron microscopy was performed by U.H. and C.A. D.B. contributed to sample preparation. M.T. and T.T. wrote the paper with support from M.S.

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Correspondence to Takashi Toda.

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The authors declare no competing financial interests.

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Supplementary Information

Supplementary figures S1, S2, S3, S4, Supplementary table S1 and Supplementary notes (PDF 0 kb)

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Toya, M., Sato, M., Haselmann, U. et al. γ-Tubulin complex-mediated anchoring of spindle microtubules to spindle-pole bodies requires Msd1 in fission yeast. Nat Cell Biol 9, 646–653 (2007). https://doi.org/10.1038/ncb1593

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