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Ran stimulates spindle assembly by altering microtubule dynamics and the balance of motor activities

Nature Cell Biology volume 3, pages 221–227 (2001)Cite this article

Abstract

The guanosine tri-phosphatase Ran stimulates assembly of microtubule spindles. However, it is not known what aspects of the microtubule cytoskeleton are subject to regulation by Ran in mitosis. Here we show that Ran–GTP stimulates microtubule assembly by increasing the rescue frequency of microtubules three- to eightfold. In addition to changing microtubule dynamics, Ran–GTP also alters the balance of motor activities, partly as a result of an increase in the amount of motile Eg5, a plus-end-directed microtubule motor that is essential for spindle formation. Thus, Ran regulates multiple processes that are involved in spindle assembly.

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Figure 1: Analysis of microtubule dynamics in CSF-arrested extracts in the presence or absence of Ran(L43E).
Figure 2: Distributions of microtubule growth and shrinkage rates.
Figure 3: Microtubule structures formed in CSF-arrested egg extracts.
Figure 4: Ran(L43E) stimulates plus-end-directed movement of microtubule seeds.
Figure 5: Microtubule seeds lead with their minus ends when moving towards the plus or minus end of the aster.
Figure 6: Ran–GTP alters the behaviour of Eg5 on microtubule asters.

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Acknowledgements

We thank T. Kapoor and T. Mitchison (Harvard Medical School) for Monastrol, O. Martin for purifying Ran and for critical reading of the manuscript, L. Cassimeris (Lehigh Univ.) for advice on analysing microtubule dynamics, E. Salmon (Univ. North Carolina) for advice on image analysis, A. Khodjakov (Wadsworth Center) for PKG cells used for centrosome isolation, and R. Gunawardane for comments on the manuscript. This work was supported by a postdoctoral fellowship from the American Cancer Society (to C.W.), NIH grants (to Y.Z. and C.E.W.), and a Pew Scholar's award (to Y.Z.).

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Author notes

  1. Andrew Wilde

    Present address: Department of Medical Genetics and Microbiology, University of Toronto, Kings College Circle, Toronto, M5S 1A8, Canada

  2. Andrew Wilde and Sofia B. Lizarraga: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution of Washington, Baltimore, 21210, Maryland , USA

    Andrew Wilde, Sofia B. Lizarraga, Lijun Zhang, Christiane Wiese & Yixian Zheng

  2. Graduate program in Biology, Johns Hopkins University , Baltimore, 21210, Maryland, USA

    Sofia B. Lizarraga & Yixian Zheng

  3. Universal Imaging Corporation, West Chester , 19380, Pennsylvania, USA

    Neal R. Gliksman

  4. Medical Sciences Program, Indiana University, Bloomington, 47405, Indiana, USA

    Claire E. Walczak

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Correspondence to Yixian Zheng.

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Wilde, A., Lizarraga, S., Zhang, L. et al. Ran stimulates spindle assembly by altering microtubule dynamics and the balance of motor activities. Nat Cell Biol 3, 221–227 (2001). https://doi.org/10.1038/35060000

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