Loss of N1-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression

Authors

Isao Masuda, Thomas Jefferson UniversityFollow
Jae-Yeon Hwang, Johns Hopkins University
Thomas Christian, Thomas Jefferson UniversityFollow
Sunita Maharjan, Thomas Jefferson UniversityFollow
Fuad Mohammad, Johns Hopkins University
Howard Gamper, Thomas Jefferson UniversityFollow
Allen R. Buskirk, Johns Hopkins University
Ya-Ming Hou, Thomas Jefferson UniversityFollow

Document Type

Article

Publication Date

8-12-2021

Comments

This article is the authors’ final published version in eLife, Volume 10, August 2021, Article number e70619.

The published version is available at https://doi.org/10.7554/eLife.70619. Copyright © Masuda et al.

Publication made possible in part by support from the Jefferson Open Access Fund

Abstract

N¹-methylation of G37 is required for a subset of tRNAs to maintain the translational reading-frame. While loss of m¹G37 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 m¹G37 deficiency on protein synthesis. Using E coli as a model, we show that m¹G37 deficiency induces ribosome stalling at codons that are normally translated by m¹G37-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 affected codons leave the A site. Enzyme- and cell-based assays show that m¹G37 deficiency reduces tRNA aminoacylation and in some cases peptide-bond formation. We observe changes of gene expression in m¹G37 deficiency similar to those in the stringent response that is typically induced by deficiency of amino acids. This work demonstrates a previously unrecognized function of m¹G37 that emphasizes its role throughout the entire elongation cycle of protein synthesis, providing new insight into its essentiality for bacterial growth and survival.

Recommended Citation

Masuda, Isao; Hwang, Jae-Yeon; Christian, Thomas; Maharjan, Sunita; Mohammad, Fuad; Gamper, Howard; Buskirk, Allen R.; and Hou, Ya-Ming, "Loss of N1-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression" (2021). Department of Biochemistry and Molecular Biology Faculty Papers. Paper 193.
https://jdc.jefferson.edu/bmpfp/193

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 License.

PubMed ID

34382933

Language

English