Bod1, a novel kinetochore protein required for chromosome biorientation.
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.
Porter
Iain M
IM
Division of Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
McClelland
Sarah E
SE
Khoudoli
Guennadi A
GA
Hunter
Christopher J
CJ
Andersen
Jens S
JS
McAinsh
Andrew D
AD
Blow
J Julian
JJ
Swedlow
Jason R
JR
D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D018385:Centrosome / Q000378:metabolism; D045584:Chromosome Positioning; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D010641:Phenotype; D010766:Phosphorylation; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D014981:Xenopus / Q000378:metabolism*; D029867:Xenopus Proteins / Q000378:metabolism*
C524692:Bod1 protein, human; D018797:Cell Cycle Proteins; C091489:KIF2C protein, human; D034741:RNA, Small Interfering; D029867:Xenopus Proteins; D016547:Kinesins