Fig. 2: m6A position in the mRNA and codon identity modulates extent of translation delay.
From: Modulation of translational decoding by m6A modification of mRNA
The coding sequences show the unmodified and first-position modified A-site AAA or ACC codon (bold). a Time courses of f[3H]Met-Val-Phe-[14C]Lys (Lys4 mRNA, open circles) and f[3H]Met-[14C]Lys (Lys2 mRNA, closed circles) formation obtained by mixing IC (0.9 μM) with the AAA (black) or m6AAA (red) with TC (0.3 μM) containing Val-tRNAVal, Phe-tRNAPhe, and [14C]Lys-tRNALys and EF-G (4 μM) or [14C]Lys-tRNALys respectively. Smooth black lines represent one-exponential fits. The reaction time τ calculated from exponential fitting is 0.08 s for AAA vs 3.3 s for m6AAA at Lys2 and 2 s for AAA vs 18 s for m6AAA at Lys4. Thus, the delay in decoding due to the modification is ~3 s at Lys2 and ~16 s for Lys4. b Dipeptide formation with Lys TC or Thr TC (0.3 μM) upon binding to IC (0.9 μM). f[3H]Met-[14C]Lys (closed circles) or f[3H]Met-[14C]Thr (open circles) dipeptides were separated by HPLC and quantified by scintillation counting. For both panels of this figure, each time course is the mean of three independent experiments with error bars representing standard deviation (N = 3). Source data are provided as a Source Data file.
