Cargando…
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...
Autores principales: | , , , , , , , |
---|---|
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 |
_version_ | 1783741910174662656 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8384417 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT masudaisao lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT hwangjaeyeon lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT christianthomas lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT maharjansunita lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT mohammadfuad lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT gamperhoward lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT buskirkallenr lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression AT houyaming lossofn1methylationofg37intrnainducesribosomestallingandreprogramsgeneexpression |