About this item:

60 Views | 42 Downloads

Author Notes:

Sohail Khoshnevis, Email: skhoshn@emory.edu OR Email: homa.ghalei@emory.edu

S.K. and H.G. designed research; S.K., R.E.D.-W., V.M., and H.G. performed research; S.K., V.M., Y.M., and H.G. contributed new reagents/analytic tools; S.K., R.E.D.-W., V.M., Y.M., and H.G. analyzed data; and S.K. and H.G. wrote the paper with input from all authors.

We thank David Bedwell, John Dinman, Alan Hinnebusch, Katrin Karbstein, and Sunnie Thompson for the gift of plasmids, strains, and antibodies. We also thank members of the H.G. laboratory, and Drs. Anita Corbett and Daniel Reines for comments on the manuscript.

The authors declare no competing interest.

Subjects:

Research Funding:

This work was supported by startup funds from Emory University and NIH Grant 1R35GM138123 (to H.G.) and by Agence nationale de la recherche Grant MetRibo2020 and Région Grand Est Fond Regional de Cooperation pour la Recherche Grant EpiARN (to Y.M.). R. E.D.-W. was supported by an NSF Graduate Research Fellowship.

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • Science & Technology - Other Topics
  • 2 '-O-methylation
  • rRNA modification
  • ribosome biogenesis
  • ribosome dynamics
  • translation regulation
  • BOX C/D
  • SACCHAROMYCES-CEREVISIAE
  • PREINITIATION COMPLEXES
  • SOLUTION CONFORMATION
  • INITIATION-FACTORS
  • STRUCTURAL BASIS
  • YEAST
  • METHYLATION
  • PROTEIN
  • FIBRILLARIN

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

Tools:

Journal Title:

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA

Volume:

Volume 119, Number 12

Publisher:

, Pages e2117334119-e2117334119

Type of Work:

Article | Final Publisher PDF

Abstract:

Protein synthesis by ribosomes is critically important for gene expression in all cells. Ribosomal RNAs (rRNAs) are marked by numerous chemical modifications. An abundant group of rRNA modifications, present in all domains of life, is 20-O-methylation guided by box C/D small nucleolar RNAs, which are part of small ribonucleoprotein complexes (snoRNPs). Although 20-O-methylations are required for the proper production of ribosomes, the mechanisms by which these modifications contribute to translation have remained elusive. Here, we show that a change in box C/D snoRNP biogenesis in actively growing yeast cells results in the production of hypo-20-O-methylated ribosomes with distinct translational properties. Using RiboMethSeq for the quantitative analysis of 20-O-methylations, we identify site-specific perturbations of the rRNA 20-O-methylation pattern and uncover sites that are not required for ribosome production under normal conditions. Characterization of the hypo-20-O-methylated ribosomes reveals significant translational fidelity defects, including frameshifting and near-cognate start codon selection. Using rRNA structural probing, we show that hypo-20-O-methylation affects the inherent dynamics of the ribosomal subunits and impacts the binding of eukaryotic translation initiation factor 1, thereby causing translational defects. Our data reveal an unforeseen spectrum of 20-O-methylation heterogeneity in yeast rRNA and suggest a significant role for rRNA 20-O-methylation in regulating cellular translation by controlling ribosome dynamics and ligand binding.

Copyright information:

© 2022 the Author(s). Published by PNAS.

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/rdf).
Export to EndNote