Figure S8: Comparison of the error load with single errors and error clusters between wt and P610L at conditions of maximal misreading (wt at 16 µM apramycin, P610L at 256 µM apramycin). Missense peptides with error clusters are less abundant in P610L than in wt cells (red circles, quantified by PRM, for the primary data please check Figure 5D). This effect depends on the distance between the first and the consecutive amino acid substitution. In contrast to error cluster formation, EF-G (P610L) has no significant effect on other misreading events: Grey circles, single errors quantified by MS1 filtering in DDA runs; black circles, single errors quantified by targeted MS (PRM, for the primary data please check Figure 5D). The type of data acquisition had no impact on this conclusion. This shows that fusA mutations prevent error cluster formation, but have otherwise no significant impact on the proteome integrity indicating that the intrinsic ribosome fidelity is unaltered, consistent with the fact that EF-G is not bound to the ribosome upon decoding. Because error frequencies are influenced by the concentrations of competing aminoacyl-tRNAs, this also suggests that stoichiometries of cellular tRNAs are not substantially altered.