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A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence

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 s...

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Autores principales: Shen, Zhou, Wang, Gang, Yang, Yiling, Shi, Jiale, Fang, Liurong, Li, Fang, Xiao, Shaobo, Fu, Zhen F., Peng, Guiqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6746460/
https://www.ncbi.nlm.nih.gov/pubmed/31350335
http://dx.doi.org/10.1074/jbc.RA119.009713
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author Shen, Zhou
Wang, Gang
Yang, Yiling
Shi, Jiale
Fang, Liurong
Li, Fang
Xiao, Shaobo
Fu, Zhen F.
Peng, Guiqing
author_facet Shen, Zhou
Wang, Gang
Yang, Yiling
Shi, Jiale
Fang, Liurong
Li, Fang
Xiao, Shaobo
Fu, Zhen F.
Peng, Guiqing
author_sort Shen, Zhou
collection PubMed
description 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.
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spelling pubmed-67464602020-03-25 A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence Shen, Zhou Wang, Gang Yang, Yiling Shi, Jiale Fang, Liurong Li, Fang Xiao, Shaobo Fu, Zhen F. Peng, Guiqing J Biol Chem Microbiology 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. American Society for Biochemistry and Molecular Biology 2019-09-13 2019-07-26 /pmc/articles/PMC6746460/ /pubmed/31350335 http://dx.doi.org/10.1074/jbc.RA119.009713 Text en © 2019 Shen et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc. This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.
spellingShingle Microbiology
Shen, Zhou
Wang, Gang
Yang, Yiling
Shi, Jiale
Fang, Liurong
Li, Fang
Xiao, Shaobo
Fu, Zhen F.
Peng, Guiqing
A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
title A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
title_full A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
title_fullStr A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
title_full_unstemmed A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
title_short A conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
title_sort conserved region of nonstructural protein 1 from alphacoronaviruses inhibits host gene expression and is critical for viral virulence
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6746460/
https://www.ncbi.nlm.nih.gov/pubmed/31350335
http://dx.doi.org/10.1074/jbc.RA119.009713
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