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  • The EMBO Journal (1999) 18, 4196 - 4209
  • doi:10.1093/emboj/18.15.4196

CENP-C is necessary but not sufficient to induce formation of a functional centromere

Tatsuo Fukagawa1, Carlos Pendon1, John Morris2 and William Brown1

  1. Biochemistry Department, University of Oxford, South Parks Road, Oxford OX1 3QU, UK
  2. Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QU, UK

Correspondence to:

William Brown, E-mail: wrab@bioch.ox.ac.uk

Received 2 March 1999; Accepted 15 June 1999; Revised 9 June 1999

CENP-C is an evolutionarily conserved centromeric protein. We have used the chicken DT40 cell line to test the idea that CENP-C is sufficient as well as necessary for the formation of a functional centromere. We have compared the effects of disrupting the localization of CENP-C with those of inducibly overexpressing the protein. Removing CENP-C from the centromere causes disassembly of the centromere protein complex and blocks cells at the metaphase–anaphase junction. Overexpressed CENP-C is associated with an increase in errors of chromosome segregation and inhibits the completion of mitosis. However, the excess CENP-C does not disrupt the native centromeres detectably and does not associate with another conserved centromere protein, ZW10. The distribution of the excess CENP-C changes during the cell cycle. In metaphase, the excess CENP-C coats the chromosome arms. At the metaphase–anaphase transition, the excess CENP-C clusters, and during interphase it is present in large bodies which form around pre-existing centromeres which are also clustered. These results indicate that CENP-C is necessary but not sufficient for the formation of a functional centromere and suggest that the structure of CENP-C may be regulated during the cell cycle.

  • Keywords:

    • centromere,
    • CENP-C,
    • DT40,
    • mitosis