Fig. 5.

Translating RNCs have a non-equilibrium delay prior to folding. a An example trajectory showing full translation of the sequence in force versus time. The blue dashed line corresponds to codon 167, when folding would begin in equilibrium conditions. τ indicates the observed delay. The inset shows a zoom to the folding region shown at higher bandwidth (267 Hz) to see the folding transitions. b Example trajectories zoomed to show just the region after passing codon 167 (blue dashed line) to the onset of folding, either the misfolded state (1st example), the C domain (2nd example), or the misfolded state with an equilibrium-like delay (3rd example). c Zoom in regions of the data in b at higher bandwidth (267 Hz) show the folding transitions. For the spike in b in the middle panel at 101.5 sec, the zoom shows that this transition is a rare event that does not correspond to N, C, or M folding as it is too small. This event does not reoccur, and was not scored as the onset of equilibrium-like folding. d The observed delay time empirical cumulative distribution function (blue dots). An exponential fit is shown as a red line, whereas the expected distribution for time until folding based on simulations using stalled RNC data is shown as a black dotted line. e Using our fit to codon position, we can also record the codon where folding first occurred (blue dots). This is compared with a simulation of equilibrium conditions (black dotted line) and a simulation assuming an additional delay step of 63 sec (red dotted line). Source data are provided as a Source Data file