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Chromosome-induced microtubule assembly mediated by TPX2 is required for spindle formation in HeLa cells

Nature Cell Biology volume 4, pages 871–879 (2002)Cite this article

Abstract

In Xenopus laevis egg extracts, TPX2 is required for the Ran-GTP-dependent assembly of microtubules around chromosomes. Here we show that interfering with the function of the human homologue of TPX2 in HeLa cells causes defects in microtubule organization during mitosis. Suppressing the expression of human TPX2 by RNA interference leads to the formation of two microtubule asters that do not interact and do not form a spindle. Our results suggest that in vivo, even in the presence of duplicated centrosomes, spindle formation requires the function of TPX2 to generate a stable bipolar spindle with overlapping antiparallel microtubule arrays. This indicates that chromosome-induced microtubule production is a general requirement for the formation of functional spindles in animal cells.

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Figure 1: Recombinant human TPX2 rescues spindle assembly in egg extracts.
Figure 2: Localization of hTPX2 in HeLa cells.
Figure 3: Localization of hTPX2 during the cell cycle in HeLa cells.
Figure 4: Overexpression of GFP–hTPX2 causes defects in microtubule organization.
Figure 5: Antibodies against hTPX2 inhibit spindle formation.
Figure 6: Suppression of hTPX2 expression by siRNA blocks spindle formation.
Figure 7: TPX2 depletion abolishes microtubule nucleation but not stabilization.
Figure 8: Chromosome-induced microtubule assembly is required for spindle assembly.

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Acknowledgements

We thank T. Zimmermann for help with confocal microscopes and image acquisition, and E. Nigg for sharing unpublished data. This work was funded by the European Molecular Biology Laboratory (EMBL). O.J.G. was supported in part by a Deutsche Forschungsgemeinschaft fellowship and in part by an EMBL postdoctoral fellowship.

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Author notes

  1. Malte Wittmann

    Present address: Zentrum für Molekulare Biologie, Im Neuenheimer Feld 282, 69120, Heidelberg, Germany

  2. Oliver J. Gruss and Malte Wittmann: These authors contributed equally to this work

Authors and Affiliations

  1. EMBL, Meyerhofstrasse 1, Heidelberg, 69 117, Germany

    Oliver J. Gruss, Malte Wittmann, Hideki Yokoyama, Rainer Pepperkok, Eric Karsenti, Iain W. Mattaj & Isabelle Vernos

  2. Department of Cell Biology, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18a, Martinsried, 82152, Germany

    Thomas Kufer & Herman Silljé

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Correspondence to Isabelle Vernos.

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Gruss, O., Wittmann, M., Yokoyama, H. et al. Chromosome-induced microtubule assembly mediated by TPX2 is required for spindle formation in HeLa cells. Nat Cell Biol 4, 871–879 (2002). https://doi.org/10.1038/ncb870

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