SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved..
Nsp3s are the largest nonstructural proteins of coronaviruses. These transmembrane proteins include papain-like proteases (PLpro) 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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:299 |
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| Enthalten in: |
The Journal of biological chemistry - 299(2023), 12 vom: 01. Dez., Seite 105346 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Yang, Mei [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 05.01.2024 Date Revised 30.01.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jbc.2023.105346 |
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| Förderinstitution / Projekttitel: |
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| 520 | |a Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a Nsp3s are the largest nonstructural proteins of coronaviruses. These transmembrane proteins include papain-like proteases (PLpro) 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a COVID | |
| 650 | 4 | |a ISG15 | |
| 650 | 4 | |a Nsp | |
| 650 | 4 | |a cell biology | |
| 650 | 4 | |a cholesterol | |
| 650 | 4 | |a deubiquitinase | |
| 650 | 4 | |a deubiquitinating enzyme | |
| 650 | 4 | |a deubiquitination | |
| 650 | 4 | |a innate immunity | |
| 650 | 4 | |a interferon | |
| 650 | 4 | |a lipid biosynthesis | |
| 650 | 4 | |a proteolysis | |
| 650 | 4 | |a ubiquitin | |
| 650 | 4 | |a virology | |
| 650 | 4 | |a virulence factor | |
| 650 | 7 | |a papain-like protease, SARS-CoV-2 |2 NLM | |
| 650 | 7 | |a EC 3.4.22.2 |2 NLM | |
| 650 | 7 | |a Peptide Hydrolases |2 NLM | |
| 650 | 7 | |a EC 3.4.- |2 NLM | |
| 650 | 7 | |a Sterol Regulatory Element Binding Protein 1 |2 NLM | |
| 650 | 7 | |a Ubiquitin |2 NLM | |
| 650 | 7 | |a INSIG1 protein, human |2 NLM | |
| 650 | 7 | |a Sterol Regulatory Element Binding Protein 2 |2 NLM | |
| 700 | 1 | |a Mariano, Jennifer |e verfasserin |4 aut | |
| 700 | 1 | |a Su, Rebecca |e verfasserin |4 aut | |
| 700 | 1 | |a Smith, Christopher E |e verfasserin |4 aut | |
| 700 | 1 | |a Das, Sudipto |e verfasserin |4 aut | |
| 700 | 1 | |a Gill, Catherine |e verfasserin |4 aut | |
| 700 | 1 | |a Andresson, Thorkell |e verfasserin |4 aut | |
| 700 | 1 | |a Loncarek, Jadranka |e verfasserin |4 aut | |
| 700 | 1 | |a Tsai, Yien Che |e verfasserin |4 aut | |
| 700 | 1 | |a Weissman, Allan M |e verfasserin |4 aut | |
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