Eg5 is static in bipolar spindles relative to tubulin: evidence for a static spindle matrix

doi:10.1083/jcb.200106011

Published 17 September 2001. doi:10.1083/jcb.200106011

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© The Rockefeller University Press, 0021-9525/2001/9/1125 $5.00
The Journal of Cell Biology, Volume 154, Number 6, September 17, 2001 1125-1134

Article

Eg5 is static in bipolar spindles relative to tubulin

: evidence for a static spindle matrix

Tarun M. Kapoor¹ and Timothy J. Mitchison²

¹ Laboratory of Chemistry and Cell Biology, Rockefeller University, New York, NY 10021
² Department of Cell Biology, Harvard Medical School, Boston, MA 02115

Address correspondence to Tarun Kapoor, Laboratory of Chemistry and Cell Biology, Rockefeller University, 1230 York Ave., Box 202, New York, NY 10021. Tel.: (212) 327-8176. Fax: (212) 327-8177. E-mail: kapoor{at}mail.rockefeller.edu

We used fluorescent speckle microscopy to probe the dynamicsof the mitotic kinesin Eg5 in Xenopus extract spindles, andcompared them to microtubule dynamics. We found significantpopulations of Eg5 that were static over several seconds whilemicrotubules flux towards spindle poles. Eg5 dynamics are frozenby adenylimidodiphosphate. Bulk turnover experiments showedthat Eg5 can exchange between the spindle and the extract witha half life of <55 s. Eg5 distribution in spindles was notperturbed by inhibition of its motor activity with monastrol,but was perturbed by inhibition of dynactin with p50 dynamitin.We interpret these data as revealing the existence of a staticspindle matrix that promotes Eg5 targeting to spindles, andtransient immobilization of Eg5 within spindles. We discussalternative interpretations of the Eg5 dynamics we observe,ideas for the biochemical nature of a spindle matrix, and implicationsfor Eg5 function.

Key Words: mitosis; Eg5; tubulin; speckle; kinesin

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