Fig. 6

The depletion of proximal out-of-frame dATGs in eukaryotic genomes. A Meta-gene analysis shows the number of genes that harbor proximal dATGs at individual positions in the yeast genome. The curves represent the local regression line (span = 2). Spearman’s correlation coefficients (ρ) and corresponding P values are shown. B–E Numbers of genes that harbor frame +1 dATGs at individual positions in the genomes of yeast (B), humans (C), Escherichia coli (D), and Bacillus subtilis (E). E. coli and B. subtilis are used as negative controls as no ribosomal scanning mechanism is required for translation initiation in prokaryotes. F The dual-luciferase reporter experiment to test the distance-dependent inhibitory effect of dAUGs in HeLa cells. The values of individual replicates are shown by black dots and the average values are shown by red lines. The firefly/Renilla activity ratios were normalized to the variant lacking additional dATG, which were 0.33, 0.74, and 0.92 for the variants with dATGs inserted at the positions +8, +14, and +20, respectively. mRNA levels were measured with quantitative PCR and normalized to the variant lacking additional dATG. P values were given by t-tests