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Published 23 June 2003. doi:10.1083/jcb.200303185
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© The Rockefeller University Press, 0021-9525/2003/6/1041 $5.00
The Journal of Cell Biology, Volume 161, Number 6, 1041-1051

Article

 The condensin complex is required for proper spindle assembly and chromosome segregation in Xenopus egg extracts

Sarah M. Wignall, Renée Deehan, Thomas J. Maresca and Rebecca Heald

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720

Address correspondence to Rebecca Heald, Dept. of Molecular and Cell Biology, 311 Life Sciences Addition, University of California, Berkeley, Berkeley, CA 94720-3200. Tel.: (510) 643-5493. Fax: (510) 643-6791. E-mail: heald{at}socrates.berkeley.edu

Chromosome condensation is required for the physical resolution and segregation of sister chromatids during cell division, but the precise role of higher order chromatin structure in mitotic chromosome functions is unclear. Here, we address the role of the major condensation machinery, the condensin complex, in spindle assembly and function in Xenopus laevis egg extracts. Immunodepletion of condensin inhibited microtubule growth and organization around chromosomes, reducing the percentage of sperm nuclei capable of forming spindles, and causing dramatic defects in anaphase chromosome segregation. Although the motor CENP-E was recruited to kinetochores pulled poleward during anaphase, the disorganized chromosome mass was not resolved. Inhibition of condensin function during anaphase also inhibited chromosome segregation, indicating its continuous requirement. Spindle assembly around DNA-coated beads in the absence of kinetochores was also impaired upon condensin inhibition. These results support an important role for condensin in establishing chromosomal architecture necessary for proper spindle assembly and chromosome segregation.

Key Words: microtubule; mitosis; kinetochore; condensation; anaphase

The online version of this article contains supplemental material.

* Abbreviations used in this paper: CSF, cytostatic factor; FRET, fluorescence resonance energy transfer; GEF, guanine nucleotide exchange factor; SMC, structural maintenance of chromosomes; topoII, topoisomerase II{alpha}; XCAP, Xenopus chromosome–associated protein.


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