Loss of N ¹ -Methylation of G37 in tRNA Induces Ribosome Stalling and Reprograms Gene Expression

N ¹ -methylation of G37 is required for a subset of tRNAs to maintain the translational reading-frame. While loss of m ¹ G37 increases ribosomal +1 frameshifting, whether it incurs additional translational defects is unknown. Here we address this question by applying ribosome profiling to gain a genome-wide view of the effects of m ¹ G37 deficiency on protein synthesis. Using E. coli as a model, we show that m ¹ G37 deficiency induces ribosome stalling at codons that are normally translated by m ¹ G37-containing tRNAs. Stalling occurs during decoding of affected codons at the ribosomal A site, indicating a distinct mechanism than that of +1 frameshifting, which occurs after the A site. Enzyme and cell-based assays show that m ¹ G37 deficiency reduces tRNA aminoacylation and in some cases peptide-bond formation. We observe changes of gene expression in m ¹ G37 deficiency similar to those in the starvation-induced stringent response, consistent with the notion that loss of tRNA aminoacylation activates the response. This work demonstrates a previously unrecognized function of m ¹ G37 that emphasizes its role throughout the entire elongation cycle of protein synthesis, providing new insight into its essentiality for bacterial growth and survival.