sui1-L96P broadly decreases discrimination against poor Kozak context of upstream AUG codons. (A) RPFs and mRNA reads on the CPA1 gene in WT and L96P cells are depicted as described in Figure 1A, with the uORF and main CDS shown in green and pink, respectively. (B) Notched box-plots showing the distributions and medians of ratios of log2 RRO values in L96P versus WT cells (ΔRROL96P) for the subset (n = 194) of 205 translated AUG_uORFs described in Figure 1F with at least 3 nt between the cap and the uORF (to allow calculation of context scores), and for the sets of 49 uORFs comprising the top (Better context) or bottom (Poor context) quartiles of these 194 AUG_uORFs ranked by AUG context score from highest to lowest. A fraction of outliers was omitted from the plots to expand the y-axis scale. (C) Notched box-plots showing the distributions and medians of AUG context scores for the same group of 194 uORFs described in B and the two sets of 49 uORFs comprising the top (Largest ΔRRO) or bottom (Smallest ΔRRO) quartiles of the 194 AUG_uORFs ranked by ΔRROL96P values high from largest to smallest. A fraction of outliers was omitted from the plots to expand the y-axis scale. (D) The sequence logo for positions −3 to −1 and +4 surrounding the AUG codons for the same groups of uORFs shown in C. The DNA rather than RNA sequence is depicted.
