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pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains

Cellular global translation is often measured using ribosome profiling or quantitative mass spectrometry, but these methods do not provide direct information at the level of elongating nascent polypeptide chains (NPCs) and associated co-translational events. Here, we describe pSNAP, a method for pro...

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Autores principales: Uchiyama, Junki, Roy, Rohini, Wang, Dan Ohtan, Morikawa, Kazuya, Kawahara, Yuka, Iwasaki, Mio, Yoshino, Chiaki, Mishima, Yuichiro, Ishihama, Yasushi, Imami, Koshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9218386/
https://www.ncbi.nlm.nih.gov/pubmed/35754732
http://dx.doi.org/10.1016/j.isci.2022.104516
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author Uchiyama, Junki
Roy, Rohini
Wang, Dan Ohtan
Morikawa, Kazuya
Kawahara, Yuka
Iwasaki, Mio
Yoshino, Chiaki
Mishima, Yuichiro
Ishihama, Yasushi
Imami, Koshi
author_facet Uchiyama, Junki
Roy, Rohini
Wang, Dan Ohtan
Morikawa, Kazuya
Kawahara, Yuka
Iwasaki, Mio
Yoshino, Chiaki
Mishima, Yuichiro
Ishihama, Yasushi
Imami, Koshi
author_sort Uchiyama, Junki
collection PubMed
description Cellular global translation is often measured using ribosome profiling or quantitative mass spectrometry, but these methods do not provide direct information at the level of elongating nascent polypeptide chains (NPCs) and associated co-translational events. Here, we describe pSNAP, a method for proteome-wide profiling of NPCs by affinity enrichment of puromycin- and stable isotope-labeled polypeptides. pSNAP does not require ribosome purification and/or chemical labeling, and captures bona fide NPCs that characteristically exhibit protein N-terminus-biased positions. We applied pSNAP to evaluate the effect of silmitasertib, a potential molecular therapy for cancer, and revealed acute translational repression through casein kinase II and mTOR pathways. We also characterized modifications on NPCs and demonstrated that the combination of different types of modifications, such as acetylation and phosphorylation in the N-terminal region of histone H1.5, can modulate interactions with ribosome-associated factors. Thus, pSNAP provides a framework for dissecting co-translational regulations on a proteome-wide scale.
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spelling pubmed-92183862022-06-24 pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains Uchiyama, Junki Roy, Rohini Wang, Dan Ohtan Morikawa, Kazuya Kawahara, Yuka Iwasaki, Mio Yoshino, Chiaki Mishima, Yuichiro Ishihama, Yasushi Imami, Koshi iScience Article Cellular global translation is often measured using ribosome profiling or quantitative mass spectrometry, but these methods do not provide direct information at the level of elongating nascent polypeptide chains (NPCs) and associated co-translational events. Here, we describe pSNAP, a method for proteome-wide profiling of NPCs by affinity enrichment of puromycin- and stable isotope-labeled polypeptides. pSNAP does not require ribosome purification and/or chemical labeling, and captures bona fide NPCs that characteristically exhibit protein N-terminus-biased positions. We applied pSNAP to evaluate the effect of silmitasertib, a potential molecular therapy for cancer, and revealed acute translational repression through casein kinase II and mTOR pathways. We also characterized modifications on NPCs and demonstrated that the combination of different types of modifications, such as acetylation and phosphorylation in the N-terminal region of histone H1.5, can modulate interactions with ribosome-associated factors. Thus, pSNAP provides a framework for dissecting co-translational regulations on a proteome-wide scale. Elsevier 2022-06-03 /pmc/articles/PMC9218386/ /pubmed/35754732 http://dx.doi.org/10.1016/j.isci.2022.104516 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Uchiyama, Junki
Roy, Rohini
Wang, Dan Ohtan
Morikawa, Kazuya
Kawahara, Yuka
Iwasaki, Mio
Yoshino, Chiaki
Mishima, Yuichiro
Ishihama, Yasushi
Imami, Koshi
pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains
title pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains
title_full pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains
title_fullStr pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains
title_full_unstemmed pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains
title_short pSNAP: Proteome-wide analysis of elongating nascent polypeptide chains
title_sort psnap: proteome-wide analysis of elongating nascent polypeptide chains
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9218386/
https://www.ncbi.nlm.nih.gov/pubmed/35754732
http://dx.doi.org/10.1016/j.isci.2022.104516
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