rRNA Nm regulate translation. Ribosomal RNA 2'-O-methylations regulate translation by impacting ribosome dynamics

Using RiboMeth-Seq for the quantitative analysis of 2’-O methylations, we identify site-specific perturbations of the rRNA 2’-O-methylation pattern and uncover sites that are not required for ribosome production under normal conditions. Characterization of the hypo 2’-O-methylated ribosomes reveals significant translational fidelity defects including frameshifting and near-cognate start codon selection. Using rRNA structural probing, we show that hypo 2’-O-methylation affects the inherent dynamics of the ribosomal subunits and impacts the binding of translation factor eIF1 thereby causing translational defects. Our data reveal an unforeseen spectrum of 2’-O-methylation heterogeneity in yeast rRNA and suggest a significant role for rRNA 2’-O-methylation in regulating cellular translation by controlling ribosome dynamics and ligand binding.