Cohesin associates with spindle poles in a mitosis-specific manner and functions in spindle assembly in vertebrate cells

Mol Biol Cell. 2009 Mar;20(5):1289-301. doi: 10.1091/mbc.e08-04-0419. Epub 2008 Dec 30.

Abstract

Cohesin is an essential protein complex required for sister chromatid cohesion. Cohesin associates with chromosomes and establishes sister chromatid cohesion during interphase. During metaphase, a small amount of cohesin remains at the chromosome-pairing domain, mainly at the centromeres, whereas the majority of cohesin resides in the cytoplasm, where its functions remain unclear. We describe the mitosis-specific recruitment of cohesin to the spindle poles through its association with centrosomes and interaction with nuclear mitotic apparatus protein (NuMA). Overexpression of NuMA enhances cohesin accumulation at spindle poles. Although transient cohesin depletion does not lead to visible impairment of normal spindle formation, recovery from nocodazole-induced spindle disruption was significantly impaired. Importantly, selective blocking of cohesin localization to centromeres, which disrupts centromeric sister chromatid cohesion, had no effect on this spindle reassembly process, clearly separating the roles of cohesin at kinetochores and spindle poles. In vitro, chromosome-independent spindle assembly using mitotic extracts was compromised by cohesin depletion, and it was rescued by addition of cohesin that was isolated from mitotic, but not S phase, cells. The combined results identify a novel spindle-associated role for human cohesin during mitosis, in addition to its function at the centromere/kinetochore regions.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Nuclear / chemistry
  • Antigens, Nuclear / metabolism
  • Cell Cycle Proteins / metabolism
  • Cell Cycle Proteins / physiology*
  • Cell Line
  • Centrosome / metabolism
  • Centrosome / ultrastructure
  • Chickens / metabolism
  • Chromatids / metabolism
  • Chromosomal Proteins, Non-Histone / metabolism
  • Chromosomal Proteins, Non-Histone / physiology*
  • Cohesins
  • HeLa Cells
  • Humans
  • Mitosis*
  • Nocodazole / pharmacology
  • Nuclear Matrix-Associated Proteins / chemistry
  • Nuclear Matrix-Associated Proteins / metabolism
  • Protein Interaction Mapping
  • Protein Structure, Tertiary
  • Spindle Apparatus / drug effects
  • Spindle Apparatus / metabolism*
  • Spindle Apparatus / ultrastructure

Substances

  • Antigens, Nuclear
  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • NUMA1 protein, human
  • Nuclear Matrix-Associated Proteins
  • Nocodazole