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Published 23 June 2003. doi:10.1083/jcb.200304144
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© The Rockefeller University Press, 0021-9525/2003/6/1021 $5.00
The Journal of Cell Biology, Volume 161, Number 6, 1021-1028

Report

A requirement for MAP kinase in the assembly and maintenance of the mitotic spindle



Melinda M. Horne and Thomas M. Guadagno

Department of Interdisciplinary Oncology, H. Lee Moffitt Comprehensive Cancer Center, and Department of Biochemistry and Molecular Biology, University of South Florida, Tampa, FL 33612

Address correspondence to Thomas M. Guadagno, H. Lee Moffitt Cancer Center & Research Institute, Building MRC 3 annex, 12902 Magnolia Drive, Tampa, FL 33612. Tel.: (813) 903-6818. Fax: (813) 903-6817. E-mail: guadagnt{at}moffitt.usf.edu

Circumstantial evidence has suggested the possibility of microtubule-associated protein (MAP) kinase's involvement in spindle regulation. To test this directly, we asked whether MAP kinase was required for spindle assembly in Xenopus egg extracts. Either the inhibition or the depletion of endogenous p42 MAP kinase resulted in defective spindle structures resembling asters or half-spindles. Likewise, an increase in the length and polymerization of microtubules was measured in aster assays suggesting a role for MAP kinase in regulating microtubule dynamics. Consistent with this, treatment of extracts with either a specific MAP kinase kinase inhibitor or a MAP kinase phosphatase resulted in the rapid disassembly of bipolar spindles into large asters. Finally, we report that mitotic progression in the absence of MAP kinase signaling led to multiple spindle abnormalities in NIH 3T3 cells. We therefore propose that MAP kinase is a key regulator of the mitotic spindle.

Key Words: MAP kinase; Rsk; spindle assembly; Xenopus; mitotic spindle

* Abbreviations used in this paper: CSF, cytostatic factor; ERK, extracellular signal-regulated kinase; MAP, microtubule-associated protein; Mek, MAP kinase kinase; MKP-1, MAP kinase phosphatase-1; Rsk, ribosomal S6 kinase; UO126, 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene.


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