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Increased missegregation and chromosome loss with decreasing chromosome size in vertebrate cells

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Abstract

Chromosome engineering has allowed the generation of an extensive and well-defined series of linear human X centromere-based minichromosomes, which has been used to investigate the influence of size and structure on chromosome segregation in vertebrate cells. A clear relationship between overall chromosome size and mitotic stability was detected, with decreasing size associated with increasing loss rates. In chicken DT40, the lower size limit for prolonged mitotic stability is approximately 550 kb: at 450 kb, there was a dramatic increase in chromosome loss, while structures of approximately 200 kb could not be recovered. In human HT1080 cells, the size threshold for mitotic stability is approximately 1.6 Mb. Minichromosomes of 0.55–1.0 Mb can be recovered, but display high loss rates. However, all minichromosomes examined exhibited more segregation errors than normal chromosomes in HT1080 cells. This error rate increases with decreased size and correlates with reduced levels of CENP-A and Aurora B. In mouse LA-9 and Indian muntjac FM7 cells, the size requirements for mitotic stability are much greater. In mouse, a human 2.7-Mb minichromosome is rarely able to propagate a kinetochore and behaves acentrically. In Indian muntjac, CENP-C associates with the human minichromosome, but the mitotic apparatus appears unable to handle its segregation.

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Acknowledgements

We are grateful to the following for their generous gifts of antibodies: Bill Earnshaw (anti-human CENP-C); Manuel Valdivia (anti-human CENP-A); Paul Kalitsis and Andy Choo (anti-mouse CENP-C and -A); Vicianne Regnier (anti-chicken CENP-A) and Tats Fukagawa (anti-chicken CENP-H and -C). For cell lines, we thank Andrew Feinberg (FM7), Jerry Shay (IndianMuntert) and Bill Colledge (CCB mouse ES cells). We thank Chris Tyler-Smith for the pGFP-PEST:IRES:Zeo reporter cassette, and Andy Jessop and Nigel Miller for their help with flow cytometry. This work was funded by project grants from the Biotechnology and Biological Sciences Research Council (BBSRC 8/GTH12583/4), the EC Framework 5 (QLK3-CT-2002-02119) and, in its early stages, by a Medical Research Council Senior Research fellowship to CJF. We thank the BBSRC for the flow cytometer research equipment initiative grant.

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  1. Kathy Mann

    Present address: Cytogenetics Department, 5th Floor, Guy's Tower, St Thomas Street, SE1 9RT, London, UK

Authors and Affiliations

  1. Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK

    Jennifer M. Spence, Walter Mills & Christine J. Farr

  2. Cell & Molecular Biology Section, Division of Biomedical Sciences, Imperial College London, South Kensington, London, SW7 2AZ, UK

    Kathy Mann & Clare Huxley

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  1. Jennifer M. Spence
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Correspondence to Christine J. Farr.

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Communicated by H. Masumoto

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Spence, J.M., Mills, W., Mann, K. et al. Increased missegregation and chromosome loss with decreasing chromosome size in vertebrate cells. Chromosoma 115, 60–74 (2006). https://doi.org/10.1007/s00412-005-0032-6

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