Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Jul 22;13:4231. doi: 10.1038/s41467-022-31852-w

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 6 — a Schematic of smFRET experiment. Movement of SSU head domain during translocation induced by addition of EF-G to immobilized PRE complexes monitored with FRET labels on ribosomal proteins S13 (S13-Cy3, green star) and L33 (L33-Cy5, red star). FRET values representing the non-swiveled (N) and swiveled (S) states are determined in independent experiments (Supplementary Fig. 9 and Table 3). b Representative smFRET time traces for SSU head movements during EF-G(wt)-induced translocation. The majority of traces (upper panel) show fluctuations between FRET 0.8 (S) and 0.5 (N) followed by a stable FRET 0.5 (N) state after translocation. 13% of traces (lower panel) show no transition to the N state in the time course of the experiment. Pie charts indicate the percentage of traces ending in N (white) or S (gray) state. Black vertical lines represent the synchronization point, i.e., the last transition to FRET 0.8 (S) (see below). c Contour plot showing distribution of FRET values representing SSU head movement during translocation on slippery mRNA by EF-G(wt). Traces were synchronized to the last transition to FRET 0.8 (S). The duration of the last FRET 0.8 state is an estimate for the duration of translocation, because back swiveling occurs simultaneously with the dissociation of EF-G from the ribosome after translocation³⁸. Histogram at the right shows distribution of FRET values after synchronization. Data are from four independent experiments (N = 4). d Representative smFRET time traces for SSU head domain movement during translocation on slippery mRNA with EF-G(Q507D). A small fraction of traces (upper panel) show fluctuations between FRET 0.8 (S) and 0.5 (N) and end in a stable FRET 0.5 (N) state after translocation. 87% of traces (lower panel) show no transition to the N state in the time course of the experiment. e Contour plot showing distribution of FRET values representing SSU head domain movement during translocation on slippery mRNA by EF-G(Q507D). Histogram at the right shows the distribution of FRET values after synchronization. Data are from three independent experiments (N = 3).