Fig. 5: smFRET spectroscopy of the ΔΨ T-box RNA.
From: Direct observation of tRNA-chaperoned folding of a dynamic mRNA ensemble
a Sequence and secondary structure of the ΔΨ T-box RNA (nts 1–77, pseudoknot deletion). The location of the inserted uridine linker between stems I and II is indicated in red. b FRET efficiency histograms from free-diffusion experiments of ΔΨ T-box RNA in the absence (top) and presence (bottom) of 10 μM tRNA. FRET efficiency values were corrected for background. c Representative binned fluorescence (upper) and FRET efficiency (lower) trajectories of immobilized ΔΨ T-box RNA in the absence of tRNA. d Upper panel: FRET efficiency histogram of immobilized ΔΨ T-box RNA in the absence of tRNA, derived from (c). FRET efficiency values were corrected for background, donor leak into the acceptor channel, γ-factor, and direct acceptor excitation (see Methods). Lower panel: 2D FRET efficiency-donor lifetime plot of immobilized ΔΨ T-box RNA without tRNA. The on-diagonal distribution indicates a lack of flexibility in ΔΨ T-box. Source data are provided as a Source Data file.
