title: Role of lysine binding residues in the global folding of the lysC riboswitch.
abstract: Riboswitches regulate gene expression by rearranging their structure upon metabolite binding. The lysine-sensing lysC riboswitch is a rare example of an RNA aptamer organized around a 5-way helical junction in which ligand binding is performed exclusively through nucleotides located at the junction core. We have probed whether the nucleotides involved in ligand binding play any role in the global folding of the riboswitch. As predicted, our findings indicate that ligand-binding residues are critical for the lysine-dependent gene regulation mechanism. We also find that these residues are not important for the establishment of key magnesium-dependent tertiary interactions, suggesting that folding and ligand recognition are uncoupled in this riboswitch for the formation of specific interactions. However, FRET assays show that lysine binding results in an additional conformational change, indicating that lysine binding may also participate in a specific folding transition. Thus, in contrast to helical junctions being primary determinants in ribozymes and rRNA folding, we speculate that the helical junction of the lysine-sensing lysC riboswitch is not employed as structural a scaffold to direct global folding, but rather has a different role in establishing RNA-ligand interactions required for riboswitch regulation. Our work suggests that helical junctions may adopt different functions such as the coordination of global architecture or the formation of specific ligand binding site.
authors.affiliation: a Department of Biology ; Faculty of Science, RNA Group, Université de Sherbrooke , Sherbrooke ; Quebec , Canada.
authors.forename: Erich
authors.initials: E
authors.lastname: Smith-Peter
authors.affiliation: a Department of Biology ; Faculty of Science, RNA Group, Université de Sherbrooke , Sherbrooke ; Quebec , Canada.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Lamontagne
authors.affiliation: a Department of Biology ; Faculty of Science, RNA Group, Université de Sherbrooke , Sherbrooke ; Quebec , Canada.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Lafontaine
mesh_terms: D052157:Aptamers, Nucleotide / Q000378:metabolism; D001483:Base Sequence; D031541:Fluorescence Resonance Energy Transfer; D007477:Ions; D008239:Lysine / Q000378:metabolism* / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D059370:RNA Folding / Q000187:drug effects; D058928:Riboswitch / Q000235:genetics*; D061806:Transcription Termination, Genetic / Q000187:drug effects
chemical_list: D052157:Aptamers, Nucleotide; D007477:Ions; D058928:Riboswitch; D008274:Magnesium; D008239:Lysine
keywords: RNA folding; Riboswitch; helical junction; lysine; metabolite-sensing