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MBC in Press, published online ahead of print March 23, 2005
Mol. Biol. Cell 10.1091/mbc.E05-02-0174

A more recent version of this article appeared on June 1, 2005
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Submitted on March 1, 2005
Accepted on March 14, 2005

Roles of Polymerization Dynamics, Opposed Motors and a Tensile Element in Governing the Length of Xenopus Extract Meiotic Spindles

T. J. Mitchison,*{dagger} P. Maddox,*{ddagger} J. Gaetz,*{sect} A. Groen,*{dagger} M. Shirasu,*{dagger} A. Desai,*|| E. D. Salmon,*{ddagger} and T. M. Kapoor*{sect}

*Marine Biological Laboratory, Woods Hole, MA 02543; {dagger}Department of Systems Biology, Harvard Medical School, Boston, MA 02115; {ddagger}Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599; {sect}Rockefeller University, New York, NY 10021; ||Department of Cellular and Molecular Medicine, University of California, San Diego, San Diego, CA 92093

Monitoring Editor: Tim Stearns

Metaphase spindles assemble to a steady state in length by mechanisms that involve microtubule dynamics and motor proteins, but are incompletely understood. We found that Xenopus extract spindles recapitulate the length of egg meiosis II spindles, using mechanisms intrinsic to the spindle. To probe these mechanisms, we perturbed microtubule polymerization dynamics and opposed motor proteins, and measured effects on spindle morphology and dynamics. Microtubules were stabilized by hexylene glycol and inhibition of the catastrophe factor MCAK (a kinesin 13, previously called XKCM) and destabilized by depolymerizing drugs. The opposed motors Eg5 and dynein were inhibited separately and together. Our results are consistent with important roles for polymerization dynamics in regulating spindle length, and for opposed motors in regulating the relative stability of bipolar versus monopolar organization. The response to microtubule destabilization suggests that an unidentified tensile element acts in parallel with these conventional factors, generating spindle shortening force.

Address correspondence to: T. J. Mitchison (timothy_mitchison{at}hms.harvard.edu)




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