Fig. 2: DMDA-PatA provides GNG motif preference on eIF4A1.

a Schematic of RNA Bind-n-Seq experiments. Randomized RNAs associated with recombinant SBP-tagged eIF4A1 in vitro were isolated and subjected to deep sequencing. b Comparison of the enrichment of 4-mer motifs with DMSO and those with DMDA-PatA. AMP-PNP and 15 pmol of recombinant eIF4A1 were included in the reaction. Motifs containing GNG are highlighted. c Box plots for motif enrichment in RNA Bind-n-Seq (with AMP-PNP) on eIF4A1 with DMSO or DMDA-PatA in the indicated 4-mer species. d Motif enrichment Z score along the titrated recombinant eIF4A1 in RNA Bind-n-Seq (with AMP-PNP) for the indicated 4-mer species with or without DMDA-PatA. The median (line) and upper/lower quartiles (shade) are shown. e, h Fluorescence polarization assay for FAM-labeled RNAs along the titrated recombinant eIF4A1 (wild type or Phe163Leu-Ile199Met mutant) with ADP and Pi. The indicated RNA sequences at 10 nM were used with or without 50 μM DMDA-PatA. The data are presented as the mean (point) and s.d. (error) for replicates (n = 3). f Rank plot for 4-mer motifs enriched in RNA Bind-n-Seq (with ADP and Pi) on eIF4A1 in the presence of DMDA-PatA. Motifs containing GNG are highlighted. g Box plots for motif enrichment in RNA Bind-n-Seq (with ADP and Pi) on eIF4A1 with DMDA-PatA in the indicated 4-mer species. In the box plots, the medians (centerlines), upper/lower quartiles (box limits), and 1.5× interquartile ranges (whiskers) are shown. The significance was calculated by the Mann‒Whitney U test (two-tailed) for all motifs (n = 256), GNG motifs (n = 31), GAG motifs (n = 8), GUG motifs (n = 8), GGG motifs (n = 8), and GCG motifs (n = 8) (c, g). Source data are provided as a Source Data file.