Fig. 5: Simultaneous measurement of tRNA-decoding fidelity with ERASE.

a, Ribosome complexes are region specifically immobilized through hybridization between a programmed tether domain of each mRNA and a surface-immobilized DNA anchor. Four complexes carrying unique A site codons are measured simultaneously: two cognate (UUU and UUC) and two near cognate (CUC and UCC). Rates of transitions between the key tRNA selection steps (IC, CR, GA and AC states) are measured by smFRET. b, FRET contour plots reveal changes in tRNA selection fidelity with and without streptomycin, a miscoding-inducing antibiotic. c, Streptomycin results in a lower rate of ternary complex binding to the ribosome for both cognate and near-cognate codons. Data points represent mean, error bars, s.e.m.; P values, two-tailed unpaired t-test. d, Log odds of forward transition through initial selection and proofreading in the absence and presence of streptomycin. Data points represent estimated log odds of state transition, error bars denote standard error of estimate from logistic regression, where P values are derived with the Wald test. Data used for c,d represent nine (no streptomycin condition) or 14 (streptomycin condition) independent repeats.