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First published online December 11, 2006
doi: 10.1242/10.1242/jcs.03277

Journal of Cell Science 119, 5057-5066 (2006)
Published by The Company of Biologists 2006
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Research Article

Essential roles for cohesin in kinetochore and spindle function in Xenopus egg extracts

Renée Deehan Kenney and Rebecca Heald*

Department of Molecular and Cell Biology, 311 Life Sciences Addition, University of California, Berkeley, CA 94720-3200, USA

* Author for correspondence (e-mail: heald{at}socrates.berkeley.edu)

Accepted 22 September 2006

To facilitate their accurate distribution by the mitotic spindle, sister chromatids are tethered during DNA replication, attached by their kinetochores and bi-oriented on the spindle, and then simultaneously released at the metaphase to anaphase transition, allowing for their segregation to opposite spindle poles. The highly conserved cohesin complex is fundamental to this process, yet its role in mitosis is not fully understood. We show that depletion of cohesin from Xenopus egg extracts impairs sister chromatid cohesion and kinetochore-microtubule interactions, causing defective spindle attachments and chromosome alignment during metaphase and mis-segregation during anaphase. In the absence of cohesin, sister kinetochore pairing and centromeric localization of chromosomal passenger proteins INCENP and aurora B were lost upon bipolar spindle attachment. However, kinetochores remained paired with normal passenger localization if bipolar spindle formation was prevented by inhibiting the kinesin-5 motor (Eg5). These observations indicate that cohesin is not required to establish sister association, but is necessary to maintain cohesion in the presence of bipolar spindle forces. Co-depletion of cohesin together with another major SMC complex, condensin, revealed cumulative effects on spindle assembly and chromosome architecture. These data underscore the essential requirement for cohesin in sister chromatid cohesion, kinetochore and spindle function.

Key words: Cohesin, Mitosis, Chromosome, Spindle, Anaphase, Chromosomal passenger




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