Tpx2, a Novel Xenopus Map Involved in Spindle Pole Organization

doi:10.1083/jcb.149.7.1405

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© The Rockefeller University Press, 0021-9525/2000//1405 $5.00
The Journal of Cell Biology, Volume 149, Number 7, , 2000 1405-1418

Original Article

Tpx2, a Novel Xenopus Map Involved in Spindle Pole Organization

Torsten Wittmann^a, Matthias Wilm^b, Eric Karsenti^a, and Isabelle Vernos^a

^a Cell Biology and Cell Biophysics Program, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany
^b Biochemical Instrumentation Program, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany
European Molecular Biology Laboratory, Cell Biology and Cell Biophysics Program, D-69117 Heidelberg, Germany.49 6221 387 51249 6221 387 342

vernos{at}embl-heidelberg.de

TPX2, the targeting protein for Xenopus kinesin-like protein2 (Xklp2), was identified as a microtubule-associated proteinthat mediates the binding of the COOH-terminal domain of Xklp2to microtubules (Wittmann, T., H. Boleti, C. Antony, E. Karsenti,and I. Vernos. 1998. J. Cell Biol. 143:673–685). Here,we report the cloning and functional characterization of XenopusTPX2. TPX2 is a novel, basic 82.4-kD protein that is phosphorylatedduring mitosis in a microtubule-dependent way. TPX2 is nuclearduring interphase and becomes localized to spindle poles inmitosis. Spindle pole localization of TPX2 requires the activityof the dynein–dynactin complex. In late anaphase TPX2becomes relocalized from the spindle poles to the midbody. TPX2is highly homologous to a human protein of unknown functionand thus defines a new family of vertebrate spindle pole components.We investigated the function of TPX2 using spindle assemblyin Xenopus egg extracts. Immunodepletion of TPX2 from mitoticegg extracts resulted in bipolar structures with disintegratingpoles and a decreased microtubule density. Addition of an excessof TPX2 to spindle assembly reactions gave rise to monopolarstructures with abnormally enlarged poles. We conclude that,in addition to its function in targeting Xklp2 to microtubuleminus ends during mitosis, TPX2 also participates in the organizationof spindle poles.

Key Words: spindle pole • microtubules • dynein– • dynactin • TPX2 • Xklp2

The online version of this article contains supplemental material.

Abbreviations used in this paper: cdc2 kinase, cyclin-dependentkinase 2; CSF, cytostatic factor; GFP, green fluorescent protein;GST, glutathione-S-transferase; KLP(s), kinesin-like protein(s);MAP(s), microtubule-associated protein(s); MAP kinase, mitogen-activatedprotein kinase; TPX2, targeting protein for Xklp2; Xklp2, Xenopuskinesin-like protein 2.

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