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Originally published In Press as doi:10.1074/jbc.M202127200 on June 26, 2002

J. Biol. Chem., Vol. 277, Issue 37, 34074-34086, September 13, 2002
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Long CTG·CAG Repeats from Myotonic Dystrophy Are Preferred Sites for Intermolecular Recombination*

Anna PluciennikDagger §, Ravi R. IyerDagger §, Marek NapieralaDagger , Jacquelynn E. LarsonDagger , Marcin Filutowicz, and Robert D. WellsDagger ||

From the Dagger  Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University System Health Science Center, Texas Medical Center, Houston, Texas 77030 and the  Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706

Homologous recombination was shown to enable the expansion of CTG·CAG repeat sequences. Other prior investigations revealed the involvement of replication and DNA repair in these genetic instabilities. Here we used a genetic assay to measure the frequency of homologous intermolecular recombination between two CTG·CAG tracts. When compared with non-repeating sequences of similar lengths, long (CTG·CAG)n repeats apparently recombine with an ~60-fold higher frequency. Sequence polymorphisms that interrupt the homogeneity of the CTG·CAG repeat tracts reduce the apparent recombination frequency as compared with the pure uninterrupted repeats. The orientation of the repeats relative to the origin of replication strongly influenced the apparent frequency of recombination. This suggests the involvement of DNA replication in the recombination process of triplet repeats. We propose that DNA polymerases stall within the CTG·CAG repeat tracts causing nicks or double-strand breaks that stimulate homologous recombination. The recombination process is RecA-dependent.

* This work was supported by Grants GM52982, NS37554, and ES11347 from the National Institutes of Health, the Robert A. Welch Foundation (to R. D. W.), and Grant GM40314 from the National Institutes of Health (to M. F.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Present address: Dept. of Biochemistry, Duke University Medical Center, 150 Nanaline H. Duke Bldg., Research Dr., Durham, NC 27710.

|| To whom correspondence should be addressed: Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University System Health Science Center, Texas Medical Center, 2121 W. Holcombe Blvd., Houston, TX 77030-3303. Tel.: 713-677-7651; Fax: 713-677-7689; E-mail: rwells@ibt.tamu.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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