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Published online October 15, 2007
doi:10.1083/jcb.200704098
The Journal of Cell Biology, Vol. 179, No. 2, 187-197
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Porter et al.
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Bod1, a novel kinetochore protein required for chromosome biorientation



Iain M. Porter1, Sarah E. McClelland3, Guennadi A. Khoudoli1, Christopher J. Hunter2, Jens S. Andersen4, Andrew D. McAinsh3, J. Julian Blow1, and Jason R. Swedlow1

1 Division of Gene Regulation and Expression and 2 Medical Research Council Protein Phosphorylation Unit, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK
3 Chromosome Segregation Laboratory, Marie Curie Research Institute, Oxted, Surrey RH8 0TL, England, UK
4 Center for Experimental Bioinformatics, University of Southern Denmark, DS-5230 Odense, Denmark

Correspondence to Jason R. Swedlow: jason{at}lifesci.dundee.ac.uk

We have combined the proteomic analysis of Xenopus laevis in vitro–assembled chromosomes with RNA interference and live cell imaging in HeLa cells to identify novel factors required for proper chromosome segregation. The first of these is Bod1, a protein conserved throughout metazoans that associates with a large macromolecular complex and localizes with kinetochores and spindle poles during mitosis. Small interfering RNA depletion of Bod1 in HeLa cells produces elongated mitotic spindles with severe biorientation defects. Bod1-depleted cells form syntelic attachments that can oscillate and generate enough force to separate sister kinetochores, suggesting that microtubule–kinetochore interactions were intact. Releasing Bod1-depleted cells from a monastrol block increases the frequency of syntelic attachments and the number of cells displaying biorientation defects. Bod1 depletion does not affect the activity or localization of Aurora B but does cause mislocalization of the microtubule depolymerase mitotic centromere- associated kinesin and prevents its efficient phosphorylation by Aurora B. Therefore, Bod1 is a novel kinetochore protein that is required for the detection or resolution of syntelic attachments in mitotic spindles.

Abbreviations used in this paper: ACA, anticentromere antibody; CENP, centromere protein; MCAK, mitotic centromere-associated kinesin; shRNA, short hairpin RNA.


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