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Originally published In Press as doi:10.1074/jbc.M202128200 on June 3, 2002

J. Biol. Chem., Vol. 277, Issue 37, 34087-34100, September 13, 2002
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Long CTG·CAG Repeat Sequences Markedly Stimulate Intramolecular Recombination*

Marek Napierala, Pawel ParniewskiDagger , Anna Pluciennik§, and Robert D. Wells

From the Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University System Health Science Center, Texas Medical Center, Houston, Texas 77030-3303 and Dagger  Center for Microbiology and Virology, Polish Academy of Sciences, 106 Lodowa Street, 93-232 Lodz, Poland

Previous studies have shown that homologous recombination is a powerful mechanism for generation of massive instabilities of the myotonic dystrophy CTG·CAG sequences. However, the frequency of recombination between the CTG·CAG tracts has not been studied. Here we performed a systematic study on the frequency of recombination between these sequences using a genetic assay based on an intramolecular plasmid system in Escherichia coli. The rate of intramolecular recombination between long CTG·CAG tracts oriented as direct repeats was extraordinarily high; recombinants were found with a frequency exceeding 12%. Recombination occurred in both RecA+ and RecA- cells but was ~2-11 times higher in the recombination proficient strain. Long CTG·CAG tracts recombined ~10 times more efficiently than non-repeating control sequences of similar length. The recombination frequency was 60-fold higher for a pair of (CTG·CAG)165 tracts compared with a pair of (CTG·CAG)17 sequences. The CTG·CAG sequences in orientation II (CTG repeats present on a lagging strand template) recombine ~2-4 times more efficiently than tracts of identical length in the opposite orientation relative to the origin of replication. This orientation effect implies the involvement of DNA replication in the intramolecular recombination between CTG·CAG sequences. Thus, long CTG·CAG tracts are hot spots for genetic recombination.

* This work was supported by Grants GM52982, NS37554, and ES11347 from the National Institutes of Health, the Robert A. Welch Foundation, and the Friedreich's Ataxia Research Alliance.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: Center for Genome Research, Institute of Biosciences and Technology, Texas A & M University, 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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