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Quality control of protein synthesis in the early elongation stage
In the early stage of bacterial translation, peptidyl-tRNAs frequently dissociate from the ribosome (pep-tRNA drop-off) and are recycled by peptidyl-tRNA hydrolase. Here, we establish a highly sensitive method for profiling of pep-tRNAs using mass spectrometry, and successfully detect a large number...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10192219/ https://www.ncbi.nlm.nih.gov/pubmed/37198183 http://dx.doi.org/10.1038/s41467-023-38077-5 |
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author | Nagao, Asuteka Nakanishi, Yui Yamaguchi, Yutaro Mishina, Yoshifumi Karoji, Minami Toya, Takafumi Fujita, Tomoya Iwasaki, Shintaro Miyauchi, Kenjyo Sakaguchi, Yuriko Suzuki, Tsutomu |
author_facet | Nagao, Asuteka Nakanishi, Yui Yamaguchi, Yutaro Mishina, Yoshifumi Karoji, Minami Toya, Takafumi Fujita, Tomoya Iwasaki, Shintaro Miyauchi, Kenjyo Sakaguchi, Yuriko Suzuki, Tsutomu |
author_sort | Nagao, Asuteka |
collection | PubMed |
description | In the early stage of bacterial translation, peptidyl-tRNAs frequently dissociate from the ribosome (pep-tRNA drop-off) and are recycled by peptidyl-tRNA hydrolase. Here, we establish a highly sensitive method for profiling of pep-tRNAs using mass spectrometry, and successfully detect a large number of nascent peptides from pep-tRNAs accumulated in Escherichia coli pth(ts) strain. Based on molecular mass analysis, we found about 20% of the peptides bear single amino-acid substitutions of the N-terminal sequences of E. coli ORFs. Detailed analysis of individual pep-tRNAs and reporter assay revealed that most of the substitutions take place at the C-terminal drop-off site and that the miscoded pep-tRNAs rarely participate in the next round of elongation but dissociate from the ribosome. These findings suggest that pep-tRNA drop-off is an active mechanism by which the ribosome rejects miscoded pep-tRNAs in the early elongation, thereby contributing to quality control of protein synthesis after peptide bond formation. |
format | Online Article Text |
id | pubmed-10192219 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-101922192023-05-19 Quality control of protein synthesis in the early elongation stage Nagao, Asuteka Nakanishi, Yui Yamaguchi, Yutaro Mishina, Yoshifumi Karoji, Minami Toya, Takafumi Fujita, Tomoya Iwasaki, Shintaro Miyauchi, Kenjyo Sakaguchi, Yuriko Suzuki, Tsutomu Nat Commun Article In the early stage of bacterial translation, peptidyl-tRNAs frequently dissociate from the ribosome (pep-tRNA drop-off) and are recycled by peptidyl-tRNA hydrolase. Here, we establish a highly sensitive method for profiling of pep-tRNAs using mass spectrometry, and successfully detect a large number of nascent peptides from pep-tRNAs accumulated in Escherichia coli pth(ts) strain. Based on molecular mass analysis, we found about 20% of the peptides bear single amino-acid substitutions of the N-terminal sequences of E. coli ORFs. Detailed analysis of individual pep-tRNAs and reporter assay revealed that most of the substitutions take place at the C-terminal drop-off site and that the miscoded pep-tRNAs rarely participate in the next round of elongation but dissociate from the ribosome. These findings suggest that pep-tRNA drop-off is an active mechanism by which the ribosome rejects miscoded pep-tRNAs in the early elongation, thereby contributing to quality control of protein synthesis after peptide bond formation. Nature Publishing Group UK 2023-05-17 /pmc/articles/PMC10192219/ /pubmed/37198183 http://dx.doi.org/10.1038/s41467-023-38077-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Nagao, Asuteka Nakanishi, Yui Yamaguchi, Yutaro Mishina, Yoshifumi Karoji, Minami Toya, Takafumi Fujita, Tomoya Iwasaki, Shintaro Miyauchi, Kenjyo Sakaguchi, Yuriko Suzuki, Tsutomu Quality control of protein synthesis in the early elongation stage |
title | Quality control of protein synthesis in the early elongation stage |
title_full | Quality control of protein synthesis in the early elongation stage |
title_fullStr | Quality control of protein synthesis in the early elongation stage |
title_full_unstemmed | Quality control of protein synthesis in the early elongation stage |
title_short | Quality control of protein synthesis in the early elongation stage |
title_sort | quality control of protein synthesis in the early elongation stage |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10192219/ https://www.ncbi.nlm.nih.gov/pubmed/37198183 http://dx.doi.org/10.1038/s41467-023-38077-5 |
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