Effects of tRNA modification on translational accuracy depend on intrinsic codon–anticodon strength

Cellular health and growth requires protein synthesis to be both efficient to ensure sufficient production, and accurate to avoid producing defective or unstable proteins. The background of misreading error frequency by individual tRNAs is as low as 2 × 10(−6) per codon but is codon-specific with some error frequencies above 10(−3) per codon. Here we test the effect on error frequency of blocking post-transcriptional modifications of the anticodon loops of four tRNAs in Escherichia coli. We find two types of responses to removing modification. Blocking modification of [Formula: see text] and [Formula: see text] increases errors, suggesting that the modifications act at least in part to maintain accuracy. Blocking even identical modifications of [Formula: see text] and [Formula: see text] has the opposite effect of decreasing errors. One explanation could be that the modifications play opposite roles in modulating misreading by the two classes of tRNAs. Given available evidence that modifications help preorder the anticodon to allow it to recognize the codons, however, the simpler explanation is that unmodified ‘weak’ tRNAs decode too inefficiently to compete against cognate tRNAs that normally decode target codons, which would reduce the frequency of misreading.