title: Translation Initiation is Controlled by RNA Folding Kinetics via a Ribosome Drafting Mechanism.
abstract: RNA folding plays an important role in controlling protein synthesis as well as other cellular processes. Existing models have focused on how RNA folding energetics control translation initiation rate under equilibrium conditions but have largely ignored the effects of nonequilibrium RNA folding. We introduce a new mechanism, called "ribosome drafting", that explains how a mRNA's folding kinetics and the ribosome's binding rate collectively control its translation initiation rate. During cycles of translation, ribosome drafting emerges whenever successive ribosomes bind to a mRNA faster than the mRNA can refold, maintaining it in a nonequilibrium state with an acceleration of protein synthesis. Using computational design, time-correlated single photon counting, and expression measurements, we demonstrate that slow-folding and fast-folding RNA structures with equivalent folding energetics can vary protein synthesis rates by 1000-fold. We determine the necessary conditions for ribosome drafting by characterizing mRNAs with rationally designed ribosome binding rates, folding kinetics, and folding energetics, confirming the predictions of a nonequilibrium Markov model of translation. Our results have widespread implications, illustrating how competitive folding and assembly kinetics can shape the gene expression machinery's sequence-structure-function relationship inside cells.
authors.affiliation: Department of Chemical Engineering and ‡Department of Biological Engineering, The Pennsylvania State University , University Park, Pennsylvania 16802, United States.
authors.forename: Amin
authors.initials: A
authors.lastname: Espah Borujeni
authors.affiliation: Department of Chemical Engineering and ‡Department of Biological Engineering, The Pennsylvania State University , University Park, Pennsylvania 16802, United States.
authors.forename: Howard M
authors.initials: HM
authors.lastname: Salis
mesh_terms: D007700:Kinetics; D008954:Models, Biological; D010442:Peptide Chain Initiation, Translational; D014176:Protein Biosynthesis / Q000235:genetics*; D059370:RNA Folding / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D012270:Ribosomes / Q000235:genetics*
chemical_list: D012333:RNA, Messenger