Figure 7.

Differential effects of uL4D mutant ribosomes on LUC reporter expression. RNA was translated in ACE prepared from wild-type Col-0 or from two independent lines of FLAG-tagged uL4D mutants. After translation of RNA for 120 min, LUC activities were measured and normalized with the Renilla luciferase (RLUC) activity of the co-translated control RNA. The corrected reporter activities (shaded box) relative to the raw reporter activity in Col-0 ACE (open box) were calculated and means ± SD of six experiments performed with two ACE batches in triplicate are shown. Asterisks indicate significant difference from raw reporter activity in Col-0 ACE (q < 0.05 by Welch's t-test with FDR correction). (A) hCMV system. 5′-hCMV:LUC(WT) and 5′-hCMV:LUC(P21A) RNAs (Supplementary Figure S10A and D) were translated. LUC activities in 5′-hCMV:LUC(WT) RNA relative to those of 5′-hCMV:LUC(P21A) RNA were calculated. (B) AtAMD1 system. 5′-AtAMD1:LUC RNA (Supplementary Figure S13A and F) was translated in the presence of 0.2 or 0.7 mM spermidine (Spd). LUC activities in the presence of 0.7 mM Spd relative to those of 0.2 mM were calculated. (C) MAGDIS system. 5′-mAMD1:LUC RNA (Supplementary Figure S14A and G) was translated in the presence of 0.2 or 0.7 mM Spd. LUC activities in the presence of 0.7 mM Spd relative to those at 0.2 mM were calculated. (D) AUG-Stop system. 5′-NIP5;1:LUC RNA (Supplementary Figure S17A and B) was translated in the presence or absence of 0.5 mM boric acid. LUC activities in the presence of boric acid relative to those in its absence were calculated.