Fig. 3: Effects of THB on the elongation cycle of translating ribosomes. | Nature Communications

Fig. 3: Effects of THB on the elongation cycle of translating ribosomes.

From: Antibiotic thermorubin tethers ribosomal subunits and impedes A-site interactions to perturb protein synthesis in bacteria

Fig. 3

a Time course of GTP hydrolysis on EF-Tu (0.3 μΜ) in ternary complex (T3) with [3H]GTP and Phe-tRNAPhe upon its delivery to the MF-mRNA-programmed 70S IC (1 μΜ) and b subsequent dipeptide formation upon tRNA accommodation and peptidyl transfer, without or with THB—pre-incubated with 70S ribosome. c Time course of f[3H]Met-Phe dipeptide formation with saturating concentration of T3 (10 μΜ) in the absence and presence of THB. GTP hydrolysis data were fitted with single-exponential function (Supplementary Eq. 38) and dipeptide data with double-exponential function (Supplementary Eq. 39). d Time course of f[3H]Met-Phe-Phe tripeptide formation on MFF mRNA-programmed 70S IC (0.5 μΜ) mixed with 10 μΜ T3 and 10 μΜ EF-G, in the absence and presence of THB. The data were fitted to a three-step sequential model (Supplementary Eq. (52)). e Comparison of time courses of di- and tripeptide formation on THB-bound ribosome. f Time evolution of dipeptide (f[3H]Met-Phe), tripeptide (f[3H]Met-Phe-Phe) and tetrapeptide (f[3H]Met-Phe-Phe-Phe) upon MFFF mRNA translation by THB-bound 70S ribosomes. The solid lines are fitting of data to a multi-step sequential model (see Supplementary Note 2 for details).

Back to article page