Cargando…

MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells

Middle East respiratory syndrome coronavirus (MERS-CoV) emerged into humans in 2012, causing highly lethal respiratory disease. The severity of disease may be, in part, because MERS-CoV is adept at antagonizing early innate immune pathways—interferon (IFN) production and signaling, protein kinase R...

Descripción completa

Detalles Bibliográficos
Autores principales: Comar, Courtney E., Otter, Clayton J., Pfannenstiel, Jessica, Doerger, Ethan, Renner, David M., Tan, Li Hui, Perlman, Stanley, Cohen, Noam A., Fehr, Anthony R., Weiss, Susan R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9173776/
https://www.ncbi.nlm.nih.gov/pubmed/35594398
http://dx.doi.org/10.1073/pnas.2123208119
_version_ 1784722092188499968
author Comar, Courtney E.
Otter, Clayton J.
Pfannenstiel, Jessica
Doerger, Ethan
Renner, David M.
Tan, Li Hui
Perlman, Stanley
Cohen, Noam A.
Fehr, Anthony R.
Weiss, Susan R.
author_facet Comar, Courtney E.
Otter, Clayton J.
Pfannenstiel, Jessica
Doerger, Ethan
Renner, David M.
Tan, Li Hui
Perlman, Stanley
Cohen, Noam A.
Fehr, Anthony R.
Weiss, Susan R.
author_sort Comar, Courtney E.
collection PubMed
description Middle East respiratory syndrome coronavirus (MERS-CoV) emerged into humans in 2012, causing highly lethal respiratory disease. The severity of disease may be, in part, because MERS-CoV is adept at antagonizing early innate immune pathways—interferon (IFN) production and signaling, protein kinase R (PKR), and oligoadenylate synthetase/ribonuclease L (OAS/RNase L)—activated in response to viral double-stranded RNA (dsRNA) generated during genome replication. This is in contrast to severe acute respiratory syndrome CoV-2 (SARS-CoV-2), which we recently reported to activate PKR and RNase L and, to some extent, IFN signaling. We previously found that MERS-CoV accessory proteins NS4a (dsRNA binding protein) and NS4b (phosphodiesterase) could weakly suppress these pathways, but ablation of each had minimal effect on virus replication. Here we investigated the antagonist effects of the conserved coronavirus endoribonuclease (EndoU), in combination with NS4a or NS4b. Inactivation of EndoU catalytic activity alone in a recombinant MERS-CoV caused little if any effect on activation of the innate immune pathways during infection. However, infection with recombinant viruses containing combined mutations with inactivation of EndoU and deletion of NS4a or inactivation of the NS4b phosphodiesterase promoted robust activation of dsRNA-induced innate immune pathways. This resulted in at least tenfold attenuation of replication in human lung–derived A549 and primary nasal cells. Furthermore, replication of these recombinant viruses could be rescued to the level of wild-type MERS-CoV by knockout of host immune mediators MAVS, PKR, or RNase L. Thus, EndoU and accessory proteins NS4a and NS4b together suppress dsRNA-induced innate immunity during MERS-CoV infection in order to optimize viral replication.
format Online
Article
Text
id pubmed-9173776
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher National Academy of Sciences
record_format MEDLINE/PubMed
spelling pubmed-91737762022-11-20 MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells Comar, Courtney E. Otter, Clayton J. Pfannenstiel, Jessica Doerger, Ethan Renner, David M. Tan, Li Hui Perlman, Stanley Cohen, Noam A. Fehr, Anthony R. Weiss, Susan R. Proc Natl Acad Sci U S A Biological Sciences Middle East respiratory syndrome coronavirus (MERS-CoV) emerged into humans in 2012, causing highly lethal respiratory disease. The severity of disease may be, in part, because MERS-CoV is adept at antagonizing early innate immune pathways—interferon (IFN) production and signaling, protein kinase R (PKR), and oligoadenylate synthetase/ribonuclease L (OAS/RNase L)—activated in response to viral double-stranded RNA (dsRNA) generated during genome replication. This is in contrast to severe acute respiratory syndrome CoV-2 (SARS-CoV-2), which we recently reported to activate PKR and RNase L and, to some extent, IFN signaling. We previously found that MERS-CoV accessory proteins NS4a (dsRNA binding protein) and NS4b (phosphodiesterase) could weakly suppress these pathways, but ablation of each had minimal effect on virus replication. Here we investigated the antagonist effects of the conserved coronavirus endoribonuclease (EndoU), in combination with NS4a or NS4b. Inactivation of EndoU catalytic activity alone in a recombinant MERS-CoV caused little if any effect on activation of the innate immune pathways during infection. However, infection with recombinant viruses containing combined mutations with inactivation of EndoU and deletion of NS4a or inactivation of the NS4b phosphodiesterase promoted robust activation of dsRNA-induced innate immune pathways. This resulted in at least tenfold attenuation of replication in human lung–derived A549 and primary nasal cells. Furthermore, replication of these recombinant viruses could be rescued to the level of wild-type MERS-CoV by knockout of host immune mediators MAVS, PKR, or RNase L. Thus, EndoU and accessory proteins NS4a and NS4b together suppress dsRNA-induced innate immunity during MERS-CoV infection in order to optimize viral replication. National Academy of Sciences 2022-05-20 2022-05-24 /pmc/articles/PMC9173776/ /pubmed/35594398 http://dx.doi.org/10.1073/pnas.2123208119 Text en Copyright © 2022 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Comar, Courtney E.
Otter, Clayton J.
Pfannenstiel, Jessica
Doerger, Ethan
Renner, David M.
Tan, Li Hui
Perlman, Stanley
Cohen, Noam A.
Fehr, Anthony R.
Weiss, Susan R.
MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
title MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
title_full MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
title_fullStr MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
title_full_unstemmed MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
title_short MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
title_sort mers-cov endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9173776/
https://www.ncbi.nlm.nih.gov/pubmed/35594398
http://dx.doi.org/10.1073/pnas.2123208119
work_keys_str_mv AT comarcourtneye merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT otterclaytonj merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT pfannenstieljessica merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT doergerethan merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT rennerdavidm merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT tanlihui merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT perlmanstanley merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT cohennoama merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT fehranthonyr merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells
AT weisssusanr merscovendoribonucleaseandaccessoryproteinsjointlyevadehostinnateimmunityduringinfectionoflungandnasalepithelialcells