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Molecular and Cellular Biology, November 2003, p. 7689-7697, Vol. 23, No. 21
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.21.7689-7697.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Human Artificial Chromosomes with Alpha Satellite-Based De Novo Centromeres Show Increased Frequency of Nondisjunction and Anaphase Lag

M. Katharine Rudd,{dagger} Robert W. Mays,{ddagger} Stuart Schwartz, and Huntington F. Willard*

Department of Genetics, Center for Human Genetics, Case Western Reserve University School of Medicine and University Hospitals of Cleveland, Cleveland, Ohio 44106

Received 14 April 2003/ Returned for modification 20 May 2003/ Accepted 15 July 2003

Human artificial chromosomes have been used to model requirements for human chromosome segregation and to explore the nature of sequences competent for centromere function. Normal human centromeres require specialized chromatin that consists of alpha satellite DNA complexed with epigenetically modified histones and centromere-specific proteins. While several types of alpha satellite DNA have been used to assemble de novo centromeres in artificial chromosome assays, the extent to which they fully recapitulate normal centromere function has not been explored. Here, we have used two kinds of alpha satellite DNA, DXZ1 (from the X chromosome) and D17Z1 (from chromosome 17), to generate human artificial chromosomes. Although artificial chromosomes are mitotically stable over many months in culture, when we examined their segregation in individual cell divisions using an anaphase assay, artificial chromosomes exhibited more segregation errors than natural human chromosomes (P < 0.001). Naturally occurring, but abnormal small ring chromosomes derived from chromosome 17 and the X chromosome also missegregate more than normal chromosomes, implicating overall chromosome size and/or structure in the fidelity of chromosome segregation. As different artificial chromosomes missegregate over a fivefold range, the data suggest that variable centromeric DNA content and/or epigenetic assembly can influence the mitotic behavior of artificial chromosomes.

* Corresponding author. Present address: Institute for Genome Sciences and Policy, Duke University, Box 3382, 103 Research Dr., Durham, NC 27710. Phone: (919) 668-4477. Fax: (919) 668-0795. E-mail: Hunt.Willard{at}duke.edu.

{dagger} Present address: Institute for Genome Sciences and Policy, Duke University, Durham, NC 27710.

{ddagger} Present address: Athersys, Inc., Cleveland, OH 44115.

Molecular and Cellular Biology, November 2003, p. 7689-7697, Vol. 23, No. 21
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.21.7689-7697.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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