Conformation of an RNA Molecule That Models the P4/P6 Junction from Group I Introns

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Abstract
We present a three-dimensional structure of a 34-nucleotide RNA molecule determined by NMR spectroscopy. The molecule was designed to form a junction between two double-helical stems whose sequence was based on the P4/P6 domain from group I introns. There are 5‘ and 3‘ single-strand overhangs at the junction of the stems. Contrary to our expectations, we found that the 3‘ end of the molecule is placed in the minor and not the major groove of the P4 helix. As a result of tertiary contacts and stacking interactions from nucleotides in the 3‘ end, the junction helices are rotated in a left-handed fashion and do not stack coaxially. This conformation is highly dependent on the presence of single-stranded nucleotides at the 3‘ overhang. When the 3‘ end is removed, the molecule assumes a radically different structure with the 5‘ end in the minor groove of the P6 helix and an overall right-handed rotation between the stems. Only one nucleotide at the 3‘ end is sufficient to change the geometry of the junction.