Folding of the lysine riboswitch: importance of peripheral elements for transcriptional regulation

• Published in: Nucleic acids research 2011-04, Vol.39 (8), p.3373-3387
• Main Authors: Blouin, Simon, Chinnappan, Raja, Lafontaine, Daniel A
• Format: Article
• Language: English
• Subjects: Aptamers, Nucleotide - chemistry; Aptamers, Nucleotide - metabolism; Bacillus subtilis; Bacillus subtilis - genetics; Base Sequence; Binding Sites; Fluorescence Resonance Energy Transfer; Gene Expression Regulation; Ligands; Lysine - metabolism; Molecular Sequence Data; Nucleic Acid Conformation; Point Mutation; Riboswitch; RNA; Transcription, Genetic
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkq1247

The Bacillus subtilis lysC lysine riboswitch modulates its own gene expression upon lysine binding through a transcription attenuation mechanism. The riboswitch aptamer is organized around a single five-way junction that provides the scaffold for two long-range tertiary interactions (loop L2-loop L3 and helix P2-loop L4)--all of this for the creation of a specific lysine binding site. We have determined that the interaction P2-L4 is particularly important for the organization of the ligand-binding site and for the riboswitch transcription attenuation control. Moreover, we have observed that a folding synergy between L2-L3 and P2-L4 allows both interactions to fold at lower magnesium ion concentrations. The P2-L4 interaction is also critical for the close juxtaposition involving stems P1 and P5. This is facilitated by the presence of lysine, suggesting an active role of the ligand in the folding transition. We also show that a previously uncharacterized stem-loop located in the expression platform is highly important for the riboswitch activity. Thus, folding elements located in the aptamer and the expression platform both influence the lysine riboswitch gene regulation.