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Published online March 3, 2008
doi:10.1083/jcb.200706189
The Journal of Cell Biology, Vol. 180, No. 5, 867-875
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Yokoyama et al.
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Report

 Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate

Hideki Yokoyama1, Oliver J. Gruss2, Sofia Rybina1, Maïwen Caudron1, Malgorzata Schelder1, Matthias Wilm1, Iain W. Mattaj1, and Eric Karsenti1

1 European Molecular Biology Laboratory, Heidelberg 69117, Germany
2 Zentrum für Molekulare Biologie der Universität Heidelberg, 69120 Heidelberg, Germany

Correspondence to Hideki Yokoyama: yokoyama{at}embl.de; or Eric Karsenti: karsenti{at}embl.de

Production of Ran–guanosine triphosphate (GTP) around chromosomes induces local nucleation and plus end stabilization of microtubules (MTs). The nuclear protein TPX2 is required for RanGTP-dependent MT nucleation. To find the MT stabilizer, we affinity purify nuclear localization signal (NLS)–containing proteins from Xenopus laevis egg extracts. This NLS protein fraction contains the MT stabilization activity. After further purification, we used mass spectrometry to identify proteins in active fractions, including cyclin-dependent kinase 11 (Cdk11). Cdk11 localizes on spindle poles and MTs in Xenopus culture cells and egg extracts. Recombinant Cdk11 demonstrates RanGTP-dependent MT stabilization activity, whereas a kinase-dead mutant does not. Inactivation of Cdk11 in egg extracts blocks RanGTP-dependent MT stabilization and dramatically decreases the spindle assembly rate. Simultaneous depletion of TPX2 completely inhibits centrosome-dependent spindle assembly. Our results indicate that Cdk11 is responsible for RanGTP-dependent MT stabilization around chromosomes and that this local stabilization is essential for normal rates of spindle assembly and spindle function.

M. Caudron's present address is Deutsches Krebsforschungszentrum B066, 69120 Heidelberg, Germany.

Abbreviations used in this paper: CSF-XB, cytostatic factor extract buffer; M-phase, metaphase; MT, microtubule.


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