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. 2018 Jul 16;46(18):9736–9748. doi: 10.1093/nar/gky636

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Elongation rates of ribosomes on slippery mRNAs with different pseudoknots sequence during the first pseudoknot-unwinding cycle. (A) Schematic drawings of ribosomal complexes from POST^K1 to POST^K1K2. (B) Typical real-time ribosome translation trace measured using Cy3-tRNA^Lys2/Cy5-L11 FRET pair. Accommodation of Cy3-tRNA^Lys2 into the A-site led to spontaneous appearance of Cy3 and FRET signals (arrow 1). Translocation from PRE^K1K2 to POST^K1K2 caused decrease of FRET accompanied by increase of Cy3 signals (arrow 2). The high FRET state between arrows 1 and 2 corresponded to PRE^K1K2 complex. (C) Schematic drawings of ribosomal complexes from POST^K1 to POST^K2. (D) Typical real-time ribosome translation trace measured using Cy3-tRNA^Lys1/Cy5-L1 FRET pair. Arriving of Cy3-tRNA^Lys1 into the E-site led to formation of the high FRET state (arrow 3). Dissociation of Cy3-tRNA^Lys1 from the E-site caused spontaneous disappearance of Cy3 and FRET signals (arrow 4). The last high FRET state between arrows 3 and 4 corresponded to POST^K1K2 complex. (E) Dwell time distributions of PRE^K1K2 complexes during ongoing elongation. (F) Plot of PRE^K1K2 dwell time versus frameshifting efficiency. (G) Dwell time distributions of POST^K1K2 complexes during ongoing elongation. (H) Plot of POST^K1K2 dwell time versus frameshifting efficiency. n is the number of events. Dwell times were extracted by single exponential fitting of dwell time distributions. Error bars were standard errors.