Bod1, a novel kinetochore protein required for chromosome biorientation

J Cell Biol. 2007 Oct 22;179(2):187-97. doi: 10.1083/jcb.200704098. Epub 2007 Oct 15.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle Proteins / metabolism*
  • Centrosome / metabolism
  • Chromosome Positioning*
  • HeLa Cells
  • Humans
  • Kinesins / metabolism
  • Kinetochores / metabolism*
  • Microtubules / metabolism
  • Phenotype
  • Phosphorylation
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Xenopus / metabolism*
  • Xenopus Proteins / metabolism*

Substances

  • Bod1 protein, human
  • Cell Cycle Proteins
  • KIF2C protein, human
  • RNA, Small Interfering
  • Xenopus Proteins
  • Kinesins