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Human CENP-H multimers colocalize with CENP-A and CENP-C at active centromere--kinetochore complexes.

Centromere and kinetochore proteins have a pivotal role in centromere structure, kinetochore formation and sister chromatid separation. However, the molecular architecture and the precise dynamic function of the centromere-kinetochore complex during mitosis remain poorly understood. Here we report the isolation and characterization of human CENP-H. Confocal microscopic analyses of HeLa cells with anti-human CENP-H-specific antibody demonstrated that CENP-H colocalizes with inner kinetochore plate proteins CENP-A and CENP-C in both interphase and metaphase. CENP-H was present outside centromeric heterochromatin, where CENP-B is localized, and inside the kinetochore corona, where CENP-E is localized during prometaphase. Furthermore, CENP-H was detected at neocentromeres, but not at inactive centromeres in stable dicentric chromosomes. In vitro binding assays of human CENP-H with centromere-kinetochore proteins suggest that the CENP-H binds to itself and MCAK, but not to CENP-A, CENP-B or CENP-C. CENP-H multimers were observed in cells in which both FLAG-tagged CENP-H and hemagglutinin-tagged CENP-H were expressed. These results suggest that CENP-H multimers localize constitutively to the inner kinetochore plate and play an important fundamental role in organization and function of the active human centromere-kinetochore complex.

Pubmed ID: 11092768


  • Sugata N
  • Li S
  • Earnshaw WC
  • Yen TJ
  • Yoda K
  • Masumoto H
  • Munekata E
  • Warburton PE
  • Todokoro K


Human molecular genetics

Publication Data

November 22, 2000

Associated Grants

  • Agency: Wellcome Trust, Id: 073915

Mesh Terms

  • Amino Acid Sequence
  • Autoantigens
  • Centromere
  • Chromosomal Proteins, Non-Histone
  • Cloning, Molecular
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • In Situ Hybridization, Fluorescence
  • Kinetochores
  • Macromolecular Substances
  • Microscopy, Fluorescence
  • Mitosis
  • Molecular Sequence Data
  • Protein Binding
  • Recombinant Fusion Proteins
  • Sequence Alignment