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Chromosoma

, Volume 106, Issue 7, pp 446–455 | Cite as

Localization of CENP-E in the fibrous corona and outer plate of mammalian kinetochores from prometaphase through anaphase

  • Carol A. Cooke
  • Bruce Schaar
  • Tim J. Yen
  • William C. Earnshaw

Abstract. 

We have conducted a detailed ultrastructural analysis of the distribution of the kinesin-related centromere protein CENP-E during mitosis in cultured human, rat kangaroo and Indian muntjac cells. Using an affinity-purified polyclonal antibody and detection by 0.8 nm colloidal gold particles, CENP-E was localized primarily to the fibrous corona of the kinetochore in prometaphase and metaphase cells. Some labeling of the kinetochore outer plate was also observed. The distribution of fibrous corona-associated CENP-E did not change dramatically following the attachment of microtubules to the kinetochore. Thus, the normal disappearance of this kinetochore substructure in conventional electron micrographs of mitotic chromosomes with attached kinetochores is not due to the corona becoming stretched along the spindle microtubules as has been suggested. Examination of cells undergoing anaphase chromatid movement revealed the presence of CENP-E still associated with the outer surface of the kinetochore plate. At the same time, the majority of detectable CENP-E in these cells was associated with the bundles of antiparallel microtubules in the central spindle. CENP-E in this region of the cell is apparently associated with the stem body matrix material. The simultaneous localization of CENP-E on centromeres and the central spindle during anaphase was confirmed by both wide-field microscopy of human cells and conventional fluorescence microscopy of rat kangaroo cells. Together, the observations reported here are consistent with models in which CENP-E has a role in promoting the poleward migration of sister chromatids during anaphase A.

Keywords

Colloidal Gold Particle Indian Muntjac Outer Plate Central Spindle Conventional Fluorescence Microscopy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • Carol A. Cooke
    • 1
  • Bruce Schaar
    • 2
  • Tim J. Yen
    • 2
  • William C. Earnshaw
    • 1
  1. 1.Institute of Cell and Molecular Biology, University of Edinburgh, Michael Swann Building, King’s Buildings, Mayfield Road, Edinburgh EH9 3JR, UKGB
  2. 2.Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, PA 19111, USAUS

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