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Control of microtubule dynamics by the antagonistic activities of XMAP215 and XKCM1 in Xenopus egg extracts

Nature Cell Biology volume 2, pages 13–19 (2000)Cite this article

Abstract

Microtubules are dynamic polymers that move stochastically between periods of growth and shrinkage, a property known as dynamic instability. Here, to investigate the mechanisms regulating microtubule dynamics in Xenopus egg extracts, we have cloned the complementary DNA encoding the microtubule-associated protein XMAP215 and investigated the function of the XMAP215 protein. Immunodepletion of XMAP215 indicated that it is a major microtubule-stabilizing factor in Xenopus egg extracts. During interphase, XMAP215 stabilizes microtubules primarily by opposing the activity of the destabilizing factor XKCM1, a member of the kinesin superfamily. These results indicate that microtubule dynamics in Xenopus egg extracts are regulated by a balance between a stabilizing factor, XMAP215, and a destabilizing factor, XKCM1.

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Figure 1: XMAP215 amino-acid sequence and homology to other members of the protein family to which it belongs.
Figure 2: Immunolocalization of XMAP215 protein in XL177 cells.
Figure 3: XMAP215 depletion from Xenopus egg extracts.
Figure 4: XMAP215 is a major regulator of microtubule dynamics.
Figure 5: Readdition of purified XMAP215 to depleted extracts.
Figure 6: Inhibition of XKCM1 partially rescues the effect of XMAP215 depletion on the catastrophe frequency.
Figure 7: Spindle formation observed in control and XMAP215-depleted extracts.

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Acknowledgements

We thank F. Senger for cell culture; D. Drechsel for help during protein purification; A. Shevchenko and A. Shevchenko for mass spectrometry analysis; A. Desai for stimulating discussions and advice; and A. Desai, I. Sassoon, P. Gönczy, K. Oegema, M. Rathman, M. Mavris and C. Gonzalez for critical reading of the manuscript. A.P. was supported by an EMBO long-term and an HSFPO long-term fellowship. K.K. was supported by an HFSPO long-term fellowship.

Correspondence and requests for materials should be addressed to A.A.H. The XMAP215 cDNA sequence has been submitted to EMBL Nucleotide Sequence Database under accession number AJ251130.

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Author notes

  1. Régis Tournebize

    Present address: Pathogénie Microbienne Moléculaire, Institut Pasteur, 28 rue du Dr Roux, 75724, Paris, Cedex 15, France

  2. Thomas U. Mayer

    Present address: Department of Cell Biology, Harvard Medical School, 250 Longwood Avenue, Boston, Massachusetts, 02115, USA

Authors and Affiliations

  1. Cell Biology Program, EMBL, Meyerhofstrasse 1, Heidelberg, 69117, Germany

    Régis Tournebize, Andrei Popov, Kazuhisa Kinoshita, Anthony J. Ashford, Sonja Rybina, Andrei Pozniakovsky, Thomas U. Mayer, Eric Karsenti & Anthony A. Hyman

  2. Medical Sciences Program, Indiana University, Jordan Hall 306, 3rd and Hawthorne, Bloomington, 47405, Indiana, USA

    Claire E. Walczak

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Tournebize, R., Popov, A., Kinoshita, K. et al. Control of microtubule dynamics by the antagonistic activities of XMAP215 and XKCM1 in Xenopus egg extracts. Nat Cell Biol 2, 13–19 (2000). https://doi.org/10.1038/71330

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