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. Author manuscript; available in PMC: 2016 Nov 5.
Published in final edited form as: Mol Cell. 2015 Oct 22;60(3):475–486. doi: 10.1016/j.molcel.2015.09.013

Figure 5. E-site tRNA dissociates more rapidly from the PRE2 complex.

Figure 5

(A) Population FRET histograms show the distribution of states in the PRE complex (first panel), POST complex (second panel), PRE2 complex after 2 min (third panel) and the PRE2 complex after 5 min (last panel). (B) Cy3-tRNAMet dissociation from the E site tracked by the loss of fluorescence via time-lapse imaging. E-site tRNA stability for 80S IC (~0.03 min−1) squares; PRE (~0.02 min−1) circles; POST (~0.02 min−1) triangles; PRE2 (~0.25 min−1) diamonds; PRE2 + eEF2 (~1.2 min−1) hexagons. Data are represented as mean +/− SD. (C) A representative pre-steady state single-molecule FRET trace of productive aa-tRNA selection showing dynamic motions within the ribosome bearing three fluorescently-labeled tRNAs. The area in green indicates the time occupied by a stable POST complex before delivery of Cy7-labeled ternary complex; tRNA selection is highlighted in blue; the PRE2 complex is highlighted in yellow. See also Figure S5.