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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 ge...

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Autores principales: Masuda, Isao, Hwang, Jae-Yeon, Christian, Thomas, Maharjan, Sunita, Mohammad, Fuad, Gamper, Howard, Buskirk, Allen R, Hou, Ya-Ming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8384417/
https://www.ncbi.nlm.nih.gov/pubmed/34382933
http://dx.doi.org/10.7554/eLife.70619
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author Masuda, Isao
Hwang, Jae-Yeon
Christian, Thomas
Maharjan, Sunita
Mohammad, Fuad
Gamper, Howard
Buskirk, Allen R
Hou, Ya-Ming
author_facet Masuda, Isao
Hwang, Jae-Yeon
Christian, Thomas
Maharjan, Sunita
Mohammad, Fuad
Gamper, Howard
Buskirk, Allen R
Hou, Ya-Ming
author_sort Masuda, Isao
collection PubMed
description 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 affected codons leave 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 stringent response that is typically induced by deficiency of amino acids. 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.
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spelling pubmed-83844172021-08-25 Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression Masuda, Isao Hwang, Jae-Yeon Christian, Thomas Maharjan, Sunita Mohammad, Fuad Gamper, Howard Buskirk, Allen R Hou, Ya-Ming eLife Microbiology and Infectious Disease 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 affected codons leave 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 stringent response that is typically induced by deficiency of amino acids. 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. eLife Sciences Publications, Ltd 2021-08-12 /pmc/articles/PMC8384417/ /pubmed/34382933 http://dx.doi.org/10.7554/eLife.70619 Text en © 2021, Masuda et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Microbiology and Infectious Disease
Masuda, Isao
Hwang, Jae-Yeon
Christian, Thomas
Maharjan, Sunita
Mohammad, Fuad
Gamper, Howard
Buskirk, Allen R
Hou, Ya-Ming
Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression
title Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression
title_full Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression
title_fullStr Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression
title_full_unstemmed Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression
title_short Loss of N(1)-methylation of G37 in tRNA induces ribosome stalling and reprograms gene expression
title_sort loss of n(1)-methylation of g37 in trna induces ribosome stalling and reprograms gene expression
topic Microbiology and Infectious Disease
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8384417/
https://www.ncbi.nlm.nih.gov/pubmed/34382933
http://dx.doi.org/10.7554/eLife.70619
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