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SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum

Nsp3s are the largest nonstructural proteins of coronaviruses. These transmembrane proteins include papain-like proteases (PLpros) that play essential roles in cleaving viral polyproteins into their mature units. The PLpro of SARS-CoV viruses also have deubiquitinating and deISGylating activities. A...

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Autores principales: Yang, Mei, Mariano, Jennifer, Su, Rebecca, Smith, Christopher E., Das, Sudipto, Gill, Catherine, Andresson, Thorkell, Loncarek, Jadranka, Tsai, Yien Che, Weissman, Allan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10692909/
https://www.ncbi.nlm.nih.gov/pubmed/37838170
http://dx.doi.org/10.1016/j.jbc.2023.105346
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author Yang, Mei
Mariano, Jennifer
Su, Rebecca
Smith, Christopher E.
Das, Sudipto
Gill, Catherine
Andresson, Thorkell
Loncarek, Jadranka
Tsai, Yien Che
Weissman, Allan M.
author_facet Yang, Mei
Mariano, Jennifer
Su, Rebecca
Smith, Christopher E.
Das, Sudipto
Gill, Catherine
Andresson, Thorkell
Loncarek, Jadranka
Tsai, Yien Che
Weissman, Allan M.
author_sort Yang, Mei
collection PubMed
description Nsp3s are the largest nonstructural proteins of coronaviruses. These transmembrane proteins include papain-like proteases (PLpros) that play essential roles in cleaving viral polyproteins into their mature units. The PLpro of SARS-CoV viruses also have deubiquitinating and deISGylating activities. As Nsp3 is an endoplasmic reticulum (ER)-localized protein, we asked if the deubiquitinating activity of SARS-CoV-2 PLpro affects proteins that are substrates for ER-associated degradation (ERAD). Using full-length Nsp3 as well as a truncated transmembrane form we interrogated, by coexpression, three potential ERAD substrates, all of which play roles in regulating lipid biosynthesis. Transmembrane PLpro increases the level of INSIG-1 and decreases its ubiquitination. However, different effects were seen with SREBP-1 and SREBP-2. Transmembrane PLpro cleaves SREBP-1 at three sites, including two noncanonical sites in the N-terminal half of the protein, resulting in a decrease in precursors of the active transcription factor. Conversely, cleavage of SREBP-2 occurs at a single canonical site that disrupts a C-terminal degron, resulting in increased SREBP-2 levels. When this site is mutated and the degron can no longer be interrupted, SREBP-2 is still stabilized by transmembrane PLpro, which correlates with a decrease in SREBP-2 ubiquitination. All of these observations are dependent on PLpro catalytic activity. Our findings demonstrate that, when anchored to the ER membrane, SARS-CoV-2 Nsp3 PLpro can function as a deubiquitinating enzyme to stabilize ERAD substrates. Additionally, SARS-CoV-2 Nsp3 PLpro can cleave ER-resident proteins, including at sites that could escape analyses based on the established consensus sequence.
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spelling pubmed-106929092023-12-03 SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum Yang, Mei Mariano, Jennifer Su, Rebecca Smith, Christopher E. Das, Sudipto Gill, Catherine Andresson, Thorkell Loncarek, Jadranka Tsai, Yien Che Weissman, Allan M. J Biol Chem Research Article Nsp3s are the largest nonstructural proteins of coronaviruses. These transmembrane proteins include papain-like proteases (PLpros) that play essential roles in cleaving viral polyproteins into their mature units. The PLpro of SARS-CoV viruses also have deubiquitinating and deISGylating activities. As Nsp3 is an endoplasmic reticulum (ER)-localized protein, we asked if the deubiquitinating activity of SARS-CoV-2 PLpro affects proteins that are substrates for ER-associated degradation (ERAD). Using full-length Nsp3 as well as a truncated transmembrane form we interrogated, by coexpression, three potential ERAD substrates, all of which play roles in regulating lipid biosynthesis. Transmembrane PLpro increases the level of INSIG-1 and decreases its ubiquitination. However, different effects were seen with SREBP-1 and SREBP-2. Transmembrane PLpro cleaves SREBP-1 at three sites, including two noncanonical sites in the N-terminal half of the protein, resulting in a decrease in precursors of the active transcription factor. Conversely, cleavage of SREBP-2 occurs at a single canonical site that disrupts a C-terminal degron, resulting in increased SREBP-2 levels. When this site is mutated and the degron can no longer be interrupted, SREBP-2 is still stabilized by transmembrane PLpro, which correlates with a decrease in SREBP-2 ubiquitination. All of these observations are dependent on PLpro catalytic activity. Our findings demonstrate that, when anchored to the ER membrane, SARS-CoV-2 Nsp3 PLpro can function as a deubiquitinating enzyme to stabilize ERAD substrates. Additionally, SARS-CoV-2 Nsp3 PLpro can cleave ER-resident proteins, including at sites that could escape analyses based on the established consensus sequence. American Society for Biochemistry and Molecular Biology 2023-10-12 /pmc/articles/PMC10692909/ /pubmed/37838170 http://dx.doi.org/10.1016/j.jbc.2023.105346 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Yang, Mei
Mariano, Jennifer
Su, Rebecca
Smith, Christopher E.
Das, Sudipto
Gill, Catherine
Andresson, Thorkell
Loncarek, Jadranka
Tsai, Yien Che
Weissman, Allan M.
SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
title SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
title_full SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
title_fullStr SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
title_full_unstemmed SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
title_short SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
title_sort sars-cov-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10692909/
https://www.ncbi.nlm.nih.gov/pubmed/37838170
http://dx.doi.org/10.1016/j.jbc.2023.105346
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