Unboxing the T‐box riboswitches—A glimpse into multivalent and multimodal RNA–RNA interactions

The T‐box riboswitches are widespread bacterial noncoding RNAs that directly bind specific tRNAs, sense aminoacylation on bound tRNAs, and switch conformations to control amino‐acid metabolism and to maintain nutritional homeostasis. The core mechanisms of tRNA recognition, amino acid sensing, and conformational switching by the T‐boxes have been recently elucidated, providing a wealth of new insights into multivalent and multimodal RNA–RNA interactions. This review dissects the structures and tRNA‐recognition mechanisms by the Stem I, Stem II, and Discriminator domains, which collectively compose the T‐box riboswitches. It further compares and contrasts the two classes of T‐boxes that regulate transcription and translation, respectively, and integrates recent findings to derive general themes, trends, and insights into complex RNA–RNA interactions. Specifically, the T‐box paradigm reveals that noncoding RNAs can interact with each other through multiple coordinated contacts, concatenation of stacked helices, and mutually induced fit. Numerous tertiary contacts, especially those emanating from strings of single‐stranded purines, act in concert to reinforce long‐range base‐pairing and stacking interactions. These coordinated, mixed‐mode contacts allow the T‐box RNA to sterically sense aminoacylation on the tRNA using a bipartite steric sieve, and to couple this readout to a conformational switch mediated by tRNA‐T‐box stacking. Together, the insights gleaned from the T‐box riboswitches inform investigations into other complex RNA structures and assemblies, development of T‐box‐targeted antimicrobials, and may inspire design and engineering of novel RNA sensors, regulators, and interfaces. This article is categorized under: RNA Structure and Dynamics > RNA Structure, Dynamics and Chemistry. Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs. Regulatory RNAs/RNAi/Riboswitches > Riboswitches.