Mechanical Forces Have a Range of Effects on the Rate of Ribosome Catalyzed Peptidyl Transfer Depending on Direction

Mechanical forces acting on the nascent chain residue located at the P-site of the ribosome can influence codon translation rates. Most observations to date involve force vectors aligned collinear with the long axis of the ribosome exit tunnel. What is poorly understood is how force applied in other directions will impact the rate of peptide bond formation catalyzed by the ribosome. Here, we utilize quantum mechanical/molecular mechanics simulations to estimate the changes in the activation free energy as a consequence of applying a constant force in various directions on the C-terminal residue at the P-site. Qualitatively consistent with the Bell model, we find this force can either accelerate, decelerate, or not alter the reaction rate depending on the force direction. A force in the average direction between the P-site 3′ O-C ester bond that breaks and the peptide bond that forms accelerates the reaction. A force in the opposite direction slows down the reaction as it opposes these bonds breaking and forming, but surprisingly it does not do so to the maximum extent possible. In this case, there is a counterbalancing trend; the force in this direction brings the A-site amino nitrogen and the P-site tRNA A76 3′ oxygen groups closer together, which promotes one of the proton shuttling steps of the reaction. We find the maximum force-induced slowdown occurs 37° off this axis. If force is applied in orthogonal directions to the reaction coordinates, there is no significant change in the reaction speed. These results indicate that there is a richer set of scenarios of force effects on translation speed that have yet to be experimentally explored and raise the possibility that cells could use these mechanochemical effects to modulate and regulate protein synthesis.