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First published online 10 February 2009
doi: 10.1242/jcs.037259

Journal of Cell Science 122, 644-655 (2009)
Published by The Company of Biologists 2009
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Research Article

Dynamic release of nuclear RanGTP triggers TPX2-dependent microtubule assembly during the apoptotic execution phase

David K. Moss1,*, Andrew Wilde2 and Jon D. Lane1,{ddagger}

1 Cell Biology Laboratories, Department of Biochemistry, University of Bristol, School of Medical Sciences, University Walk, Bristol BS8 1TD, UK
2 Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada

{ddagger} Author for correspondence (e-mail: jon.lane{at}bristol.ac.uk)

Accepted 3 November 2008

During apoptosis, the interphase microtubule network is dismantled then later replaced by a novel, non-centrosomal microtubule array. These microtubules assist in the peripheral redistribution of nuclear fragments in the apoptotic cell; however, the regulation of apoptotic microtubule assembly is not understood. Here, we demonstrate that microtubule assembly depends upon the release of nuclear RanGTP into the apoptotic cytoplasm because this process is blocked in apoptotic cells overexpressing dominant-negative GDP-locked Ran (T24N). Actin–myosin-II contractility provides the impetus for Ran release and, consequently, microtubule assembly is blocked in blebbistatin- and Y27632-treated apoptotic cells. Importantly, the spindle-assembly factor TPX2 (targeting protein for Xklp2), colocalises with apoptotic microtubules, and siRNA silencing of TPX2, but not of the microtubule motors Mklp1 and Kid, abrogates apoptotic microtubule assembly. These data provide a molecular explanation for the assembly of the apoptotic microtubule network, and suggest important similarities with the process of RanGTP- and TPX2-mediated mitotic spindle formation.

Key words: Apoptosis, Microtubules, Ran, TPX2


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