A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
© 2019 Shen et al..
Coronaviruses are enveloped, single-stranded RNA viruses that are distributed worldwide. They include transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), and the human coronaviruses severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV), many of which seriously endanger human health and well-being. Only alphacoronaviruses and betacoronaviruses harbor nonstructural protein 1 (nsp1), which performs multiple functions in inhibiting antiviral host responses. The role of the C terminus of betacoronavirus nsp1 in virulence has been characterized, but the location of the alphacoronavirus nsp1 region that is important for virulence remains unclear. Here, using TGEV nsp1 as a model to explore the function of this protein in alphacoronaviruses, we demonstrate that alphacoronavirus nsp1 inhibits host gene expression. Solving the crystal structure of full-length TGEV at 1.85-Å resolution and conducting several biochemical analyses, we observed that a specific motif (amino acids 91-95) of alphacoronavirus nsp1 is a conserved region that inhibits host protein synthesis. Using a reverse-genetics system based on CRISPR/Cas9 technology to construct a recombinant TGEV in which this specific nsp1 motif was altered, we found that this mutation does not affect virus replication in cell culture but significantly reduces TGEV pathogenicity in pigs. Taken together, our findings suggest that alphacoronavirus nsp1 is an essential virulence determinant, providing a potential paradigm for the development of a new attenuated vaccine based on modified nsp1.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2019 |
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| Erschienen: |
2019 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:294 |
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| Enthalten in: |
The Journal of biological chemistry - 294(2019), 37 vom: 13. Sept., Seite 13606-13618 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Shen, Zhou [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 12.05.2020 Date Revised 07.12.2022 published: Print-Electronic PDB: 3ZBD, 6IVC, 6IVD Citation Status MEDLINE |
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| doi: |
10.1074/jbc.RA119.009713 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM299620557 |
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| 100 | 1 | |a Shen, Zhou |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence |
| 264 | 1 | |c 2019 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
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| 500 | |a Date Completed 12.05.2020 | ||
| 500 | |a Date Revised 07.12.2022 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a PDB: 3ZBD, 6IVC, 6IVD | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2019 Shen et al. | ||
| 520 | |a Coronaviruses are enveloped, single-stranded RNA viruses that are distributed worldwide. They include transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), and the human coronaviruses severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV), many of which seriously endanger human health and well-being. Only alphacoronaviruses and betacoronaviruses harbor nonstructural protein 1 (nsp1), which performs multiple functions in inhibiting antiviral host responses. The role of the C terminus of betacoronavirus nsp1 in virulence has been characterized, but the location of the alphacoronavirus nsp1 region that is important for virulence remains unclear. Here, using TGEV nsp1 as a model to explore the function of this protein in alphacoronaviruses, we demonstrate that alphacoronavirus nsp1 inhibits host gene expression. Solving the crystal structure of full-length TGEV at 1.85-Å resolution and conducting several biochemical analyses, we observed that a specific motif (amino acids 91-95) of alphacoronavirus nsp1 is a conserved region that inhibits host protein synthesis. Using a reverse-genetics system based on CRISPR/Cas9 technology to construct a recombinant TGEV in which this specific nsp1 motif was altered, we found that this mutation does not affect virus replication in cell culture but significantly reduces TGEV pathogenicity in pigs. Taken together, our findings suggest that alphacoronavirus nsp1 is an essential virulence determinant, providing a potential paradigm for the development of a new attenuated vaccine based on modified nsp1 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a coronavirus | |
| 650 | 4 | |a crystal structure | |
| 650 | 4 | |a host gene expression | |
| 650 | 4 | |a immune evasion | |
| 650 | 4 | |a nonstructural protein 1 (nsp1) | |
| 650 | 4 | |a pathogenesis | |
| 650 | 4 | |a protein motif | |
| 650 | 4 | |a transmissible gastroenteritis virus (TGEV) | |
| 650 | 4 | |a virulence factor | |
| 650 | 4 | |a virus | |
| 650 | 7 | |a Synapsins |2 NLM | |
| 650 | 7 | |a Viral Nonstructural Proteins |2 NLM | |
| 650 | 7 | |a Nsp1 protein, SARS coronavirus |2 NLM | |
| 650 | 7 | |a EC 2.7.7.48 |2 NLM | |
| 650 | 7 | |a RNA-Dependent RNA Polymerase |2 NLM | |
| 650 | 7 | |a EC 2.7.7.48 |2 NLM | |
| 700 | 1 | |a Wang, Gang |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Yiling |e verfasserin |4 aut | |
| 700 | 1 | |a Shi, Jiale |e verfasserin |4 aut | |
| 700 | 1 | |a Fang, Liurong |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Fang |e verfasserin |4 aut | |
| 700 | 1 | |a Xiao, Shaobo |e verfasserin |4 aut | |
| 700 | 1 | |a Fu, Zhen F |e verfasserin |4 aut | |
| 700 | 1 | |a Peng, Guiqing |e verfasserin |4 aut | |
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