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Nucleic Acids Research 2005 33(5):1604-1617; doi:10.1093/nar/gki307
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Published online 14 March 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
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Article

Structural roles of CTG repeats in slippage expansion during DNA replication

Lai Man Chi and Sik Lok Lam*

Department of Chemistry, The Chinese University of Hong Kong Shatin, New Territories, Hong Kong

*To whom correspondence should be addressed. Tel: +852 2609 8126; Fax: +852 2603 5057; Email: lams{at}cuhk.edu.hk

Received November 25, 2004. Revised December 29, 2004. Accepted February 27, 2005.

CTG triplet repeat sequences have been found to form slipped-strand structures leading to self-expansion during DNA replication. The lengthening of these repeats causes the onset of neurodegenerative diseases, such as myotonic dystrophy. In this study, electrophoretic and NMR spectroscopic studies have been carried out to investigate the length and the structural roles of CTG repeats in affecting the hairpin formation propensity. Direct NMR evidence has been successfully obtained the first time to support the presence of three types of hairpin structures in sequences containing 1–10 CTG repeats. The first type contains no intra-loop hydrogen bond and occurs when the number of repeats is less than four. The second type has a 4 nt TGCT-loop and occurs in sequences with even number of repeats. The third type contains a 3 nt CTG-loop and occurs in sequences with odd number of repeats. Although stabilizing interactions have been identified between CTG repeats in both the second and third types of hairpins, the structural differences observed account for the higher hairpin formation propensity in sequences containing even number of CTG repeats. The results of this study confirm the hairpin loop structures and explain how slippage occurs during DNA replication.


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