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Authors: Marta Zumwalt, Anna Ludwig, Paul J. Hagerman and Thorsten Dieckmann

Marta Zumwalt

Department of Chemistry, University of California at Davis, Davis, California USA

Anna Ludwig

Department of Biochemistry and Molecular Medicine, University of California at Davis, Davis, California USA

Paul J. Hagerman

Department of Biochemistry and Molecular Medicine, University of California at Davis, Davis, California USA

Thorsten Dieckmann

Department of Chemistry, University of Waterloo, Waterloo, ON, Canada

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Abstract:

The CGG triplet repeat found within the 5’UTR of the FMR1 gene is involved in the pathogenesis of both fragile X syndrome and fragile X-associated tremor/ataxia syndrome (FXTAS). The repeat has been shown to form both hairpins and tetraplexes in DNA; however, the secondary structure of CGG-repeat RNA has not been well defined. To this end, we have performed NMR spectroscopy on in vitro transcribed CGG-repeat RNAs and see clear evidence of intramolecular hairpins, with no evidence of tetraplex structures. Both C•G and G◦G base pairs form in the hairpin stem, though in a dynamic equilibrium of conformations. In addition, we investigated the effect of an AGG repeat interruption on hairpin stability; such interruptions are often interspersed within the CGG repeat element and are thought to modulate secondary structure of the RNA. While the AGG repeat lowers the Tm of the hairpin at low Mg2+ concentrations, this difference disappears at physiological Mg2+ levels.