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Crystal Structure of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune Suppression
Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly emerging human pathogen that was first isolated in 2012. MERS-CoV replication depends in part on a virus-encoded papain-like protease (PL(pro)) that cleaves the viral replicase polyproteins at three sites releasing non-structural pro...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular
Biology
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4263872/ https://www.ncbi.nlm.nih.gov/pubmed/25320088 http://dx.doi.org/10.1074/jbc.M114.609644 |
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author | Bailey-Elkin, Ben A. Knaap, Robert C. M. Johnson, Garrett G. Dalebout, Tim J. Ninaber, Dennis K. van Kasteren, Puck B. Bredenbeek, Peter J. Snijder, Eric J. Kikkert, Marjolein Mark, Brian L. |
author_facet | Bailey-Elkin, Ben A. Knaap, Robert C. M. Johnson, Garrett G. Dalebout, Tim J. Ninaber, Dennis K. van Kasteren, Puck B. Bredenbeek, Peter J. Snijder, Eric J. Kikkert, Marjolein Mark, Brian L. |
author_sort | Bailey-Elkin, Ben A. |
collection | PubMed |
description | Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly emerging human pathogen that was first isolated in 2012. MERS-CoV replication depends in part on a virus-encoded papain-like protease (PL(pro)) that cleaves the viral replicase polyproteins at three sites releasing non-structural protein 1 (nsp1), nsp2, and nsp3. In addition to this replicative function, MERS-CoV PL(pro) was recently shown to be a deubiquitinating enzyme (DUB) and to possess deISGylating activity, as previously reported for other coronaviral PL(pro) domains, including that of severe acute respiratory syndrome coronavirus. These activities have been suggested to suppress host antiviral responses during infection. To understand the molecular basis for ubiquitin (Ub) recognition and deconjugation by MERS-CoV PL(pro), we determined its crystal structure in complex with Ub. Guided by this structure, mutations were introduced into PL(pro) to specifically disrupt Ub binding without affecting viral polyprotein cleavage, as determined using an in trans nsp3↓4 cleavage assay. Having developed a strategy to selectively disable PL(pro) DUB activity, we were able to specifically examine the effects of this activity on the innate immune response. Whereas the wild-type PL(pro) domain was found to suppress IFN-β promoter activation, PL(pro) variants specifically lacking DUB activity were no longer able to do so. These findings directly implicate the DUB function of PL(pro), and not its proteolytic activity per se, in the inhibition of IFN-β promoter activity. The ability to decouple the DUB activity of PL(pro) from its role in viral polyprotein processing now provides an approach to further dissect the role(s) of PL(pro) as a viral DUB during MERS-CoV infection. |
format | Online Article Text |
id | pubmed-4263872 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American Society for Biochemistry and Molecular
Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-42638722015-12-12 Crystal Structure of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune Suppression Bailey-Elkin, Ben A. Knaap, Robert C. M. Johnson, Garrett G. Dalebout, Tim J. Ninaber, Dennis K. van Kasteren, Puck B. Bredenbeek, Peter J. Snijder, Eric J. Kikkert, Marjolein Mark, Brian L. J Biol Chem Microbiology Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly emerging human pathogen that was first isolated in 2012. MERS-CoV replication depends in part on a virus-encoded papain-like protease (PL(pro)) that cleaves the viral replicase polyproteins at three sites releasing non-structural protein 1 (nsp1), nsp2, and nsp3. In addition to this replicative function, MERS-CoV PL(pro) was recently shown to be a deubiquitinating enzyme (DUB) and to possess deISGylating activity, as previously reported for other coronaviral PL(pro) domains, including that of severe acute respiratory syndrome coronavirus. These activities have been suggested to suppress host antiviral responses during infection. To understand the molecular basis for ubiquitin (Ub) recognition and deconjugation by MERS-CoV PL(pro), we determined its crystal structure in complex with Ub. Guided by this structure, mutations were introduced into PL(pro) to specifically disrupt Ub binding without affecting viral polyprotein cleavage, as determined using an in trans nsp3↓4 cleavage assay. Having developed a strategy to selectively disable PL(pro) DUB activity, we were able to specifically examine the effects of this activity on the innate immune response. Whereas the wild-type PL(pro) domain was found to suppress IFN-β promoter activation, PL(pro) variants specifically lacking DUB activity were no longer able to do so. These findings directly implicate the DUB function of PL(pro), and not its proteolytic activity per se, in the inhibition of IFN-β promoter activity. The ability to decouple the DUB activity of PL(pro) from its role in viral polyprotein processing now provides an approach to further dissect the role(s) of PL(pro) as a viral DUB during MERS-CoV infection. American Society for Biochemistry and Molecular Biology 2014-12-12 2014-10-15 /pmc/articles/PMC4263872/ /pubmed/25320088 http://dx.doi.org/10.1074/jbc.M114.609644 Text en © 2014 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 Bailey-Elkin, Ben A. Knaap, Robert C. M. Johnson, Garrett G. Dalebout, Tim J. Ninaber, Dennis K. van Kasteren, Puck B. Bredenbeek, Peter J. Snijder, Eric J. Kikkert, Marjolein Mark, Brian L. Crystal Structure of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune Suppression |
title | Crystal Structure of the Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted
Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune
Suppression |
title_full | Crystal Structure of the Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted
Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune
Suppression |
title_fullStr | Crystal Structure of the Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted
Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune
Suppression |
title_full_unstemmed | Crystal Structure of the Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted
Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune
Suppression |
title_short | Crystal Structure of the Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) Papain-like Protease Bound to Ubiquitin Facilitates Targeted
Disruption of Deubiquitinating Activity to Demonstrate Its Role in Innate Immune
Suppression |
title_sort | crystal structure of the middle east respiratory syndrome coronavirus
(mers-cov) papain-like protease bound to ubiquitin facilitates targeted
disruption of deubiquitinating activity to demonstrate its role in innate immune
suppression |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4263872/ https://www.ncbi.nlm.nih.gov/pubmed/25320088 http://dx.doi.org/10.1074/jbc.M114.609644 |
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