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. 2008 May 22;27(12):1706–1717. doi: 10.1038/emboj.2008.99

Figure 5.

Figure 5

Effects of IF3, Mg2+, mRNA and tRNA identity on the ribosomal splitting by RRF/EF-G and IF1/IF3. (A) Mixture 1 containing 0.2 μM vacant 70S ribosomes (▵,▴) or 0.2 μM 70S ribosomes pre-incubated with 0.5 μM mXR7 mRNA and 0.4 μM deacylated tRNAPhe (○,•,▪) was rapidly mixed with mixture 2 containing 9 μM RRF, 4 μM EF-G and no IF3 (open symbols) or 0.3 μM IF3 (closed symbols). (B) Mixture 1 containing 0.2 μM 70S ribosomes pre-incubated with 0.5 μM of mXR7 (•,○) or mBar (♦,⋄) mRNAs together with 0.4 μM tRNAPhe (closed symbols) or 0.4 μM tRNAfMet (open symbols) was rapidly mixed with mixture 2 containing 9 μM RRF, 4 μM EF-G and 0.3 μM IF3 or with mixture 2 containing 15 μM IF1 and 2 μM IF3. (C) Mixture 1 containing 0.2 μM vacant 70S ribosomes (▴), 0.2 μM 70S ribosomes pre-incubated with 0.5 μM of mSD022 (□), mBar (⧫), mXR8_FM (▪), mXR8 (◊) or mXR7 (•) mRNAs was rapidly mixed with mixture 2 containing 5 μM IF1 and 2 μM IF3. All mixtures were prepared in LS3 buffer, except that experiment marked (▪) in (A) was conducted in LS5 buffer containing 5 mM free Mg2+.