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Published online 18 February 2004

Nucleic Acids Research, 2004, Vol. 32, No. 3 1177-1183
© 2004 Oxford University Press

Identification of NH...N hydrogen bonds by magic angle spinning solid state NMR in a double-stranded RNA associated with myotonic dystrophy

Jörg Leppert, Carl R. Urbinati1, Sabine Häfner, Oliver Ohlenschläger, Maurice S. Swanson1, Matthias Görlach and Ramadurai Ramachandran*

Abteilung Molekulare Biophysik/NMR-Spektroskopie, Institut für Molekulare Biotechnologie, 07745 Jena, Germany and 1 Department of Molecular Genetics and Microbiology, Powell Gene Therapy Center, University of Florida, College of Medicine, 1600 SW Archer Road, Gainesville, FL 32610-0266, USA

*To whom correspondence should be addressed. Tel: +49 3641 656222 or +49 3641 656220; Fax: +49 3641 656225; Email: raman{at}imb-jena.de Correspondence may also be addressed to Matthias Görlach. Email: mago{at}imb-jena.de

RNA plays a central role in biological processes and exhibits a variety of secondary and tertiary structural features that are often stabilized via hydrogen bonds. The distance between the donor and acceptor nitrogen nuclei involved in NH...N hydrogen bonds in nucleic acid base pairs is typically in the range of 2.6–2.9 Å. Here, we show for the first time that such spatial proximity between 15N nitrogen nuclei can be conveniently monitored via magic angle spinning solid state NMR on a uniformly 15N-labelled RNA. The presence of NH...N hydrogen bonds is reflected as cross-peaks between the donor and acceptor nitrogen nuclei in 2D 15N dipolar chemical shift correlation spectra. The RNA selected for this experimental study was a CUG repeat expansion implicated in the neuromuscular disease myotonic dystrophy. The results presented provide direct evidence that the CUG repeat expansion adopts a double-stranded conformation.


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