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Evidence of cis-acting factors in replication-mediated trinucleotide repeat instability in primate cells

Nature Genetics volume 31, pages 37–46 (2002)Cite this article

Abstract

The mechanism of disease-associated trinucleotide repeat instability involves cis-acting factors (cis-elements) in the vicinity of the repeat, but the nature of these elements is unknown. One cis-element may be the location of the replication origin relative to the repeat. We have used an SV40 DNA replication system to investigate the effect of the location of replication initiation on (CTG)n•(CAG)n stability in primate cells. Depending on the distance between the SV40 replication origin and the repeat tract, templates with 79 repeats yield predominantly expansions or predominantly deletions or remain intact. All templates with 17 repeats are stable. Thus, cis-elements that affect the sites of Okazaki fragment initiation relative to the repeat are crucial determinants of instability. This model system recapitulates the bias for expansions observed in many of the diseases associated with trinucleotide repeats. Our results might explain the variable amounts of CTG/CAG instability that are observed in different chromosomal contexts.

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Figure 1: Replication fork and replication templates.
Figure 2: Experimental strategy.
Figure 3: Representative example of STRIP analysis.
Figure 4: Mutation analysis.
Figure 5: Independent analyses of primate cell–replicated material.
Figure 6: Replication fork dynamics and instability of CTG and CAG repeats.

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Acknowledgements

We thank our colleagues at The Hospital for Sick Children for comments and support, and A. Paterson for statistical analysis. This work was supported by grants from the Muscular Dystrophy Association USA and the Canadian Institutes of Health Research (CIHR) to C.E.P. and from the NIH and the March of Dimes to Y.-H.W. J.D.C. is supported by Natural Sciences and Engineering Research Council of Canada. C.E.P. is a CIHR Scholar, a Canadian Genetic Disease Network Scholar and a Premier's Research Excellence Award scholar. Y.-H.W. is a Basil O'Connor March of Dimes Fellow.

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Authors and Affiliations

  1. Program of Genetics & Genomic Biology, The Hospital for Sick Children, 555 University Avenue, Elm Wing 11-135, Toronto, M5G 1X8, Ontario, Canada

    John D. Cleary, Kerrie Nichol & Christopher E. Pearson

  2. Department of Molecular & Medical Genetics, University of Toronto,

    John D. Cleary & Christopher E. Pearson

  3. Department of Biochemistry, University of Medicine & Dentistry of New Jersey Robert Wood Johnson Medical School, Piscataway, New Jersey, USA

    Yuh-Hwa Wang

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Correspondence to Christopher E. Pearson.

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Cleary, J., Nichol, K., Wang, YH. et al. Evidence of cis-acting factors in replication-mediated trinucleotide repeat instability in primate cells. Nat Genet 31, 37–46 (2002). https://doi.org/10.1038/ng870

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