TY - JOUR
T1 - SARS-CoV-2 proteases PLpro and 3CLpro cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1)
T2 - implications for disease presentation across species
AU - Moustaqil, Mehdi
AU - Ollivier, Emma
AU - Chiu, Hsin Ping
AU - Van Tol, Sarah
AU - Rudolffi-Soto, Paulina
AU - Stevens, Christian
AU - Bhumkar, Akshay
AU - Hunter, Dominic J.B.
AU - Freiberg, Alexander N.
AU - Jacques, David
AU - Lee, Benhur
AU - Sierecki, Emma
AU - Gambin, Yann
N1 - Funding Information:
The authors would like to thank Katherina Michie, Jack Bennett and key personnel at the protein production facility of UNSW for the purification of PLpro and 3CLpro of SARS-CoV-2. The authors would like to thank Prof Michelle Baker and Claire Dickson for useful discussions and comments. The authors would like to thank Dr Kate Schroeder for the gift of the NLRP12 mouse plasmid in a previous collaboration, and Prof. Alexandrov for the cell-free plasmids compatible with LTE protein production. BL and AF acknowledge support from NIH grants AI123449 and AI125536. BL also acknowledges the Ward-Coleman estate for endowing the Ward-Coleman Chairs at the Icahn School of Medicine at Mount Sinai (ISMMS), and for additional funding support from the Department of Microbiology at ISMMS. HPC was supported by a MoST Postdoctoral Research Abroad Program 110-2917-I-564-020 (Ministry of Science and Technology, Taiwan). MM and EO performed data collection for cell-free HIIPs cleavage experiments; H-PC performed SARS-CoV-2 infection experiments, PRS performed data collection of cell-free HIIPs, AB and DH contributed to the manufacture of the in-house LTE reagents and supplementation of the extracts, AF supervised collection of SARS-CoV-2 infection data and contributed to design of the experiments. DJ designed the constructs for NSP3 and NSP5, supervised their purification and wrote the manuscript. BL contributed to the design of the experiments, supervised the data collection on SARS-CoV-2 infected cells, and wrote the manuscript. ES and YG designed the project, supervised cell-free experiments on HIIPs and wrote the manuscript.
Funding Information:
The authors would like to thank Katherina Michie, Jack Bennett and key personnel at the protein production facility of UNSW for the purification of PLpro and 3CLpro of SARS-CoV-2. The authors would like to thank Prof Michelle Baker and Claire Dickson for useful discussions and comments. The authors would like to thank Dr Kate Schroeder for the gift of the NLRP12 mouse plasmid in a previous collaboration, and Prof. Alexandrov for the cell-free plasmids compatible with LTE protein production. BL and AF acknowledge support from NIH grants AI123449 and AI125536. BL also acknowledges the Ward-Coleman estate for endowing the Ward-Coleman Chairs at the Icahn School of Medicine at Mount Sinai (ISMMS), and for additional funding support from the Department of Microbiology at ISMMS. HPC was supported by a MoST Postdoctoral Research Abroad Program 110-2917-I-564-020 (Ministry of Science and Technology, Taiwan).
Publisher Copyright:
© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd.
PY - 2021
Y1 - 2021
N2 - The genome of SARS-CoV-2 encodes two viral proteases (NSP3/papain-like protease and NSP5/3C-like protease) that are responsible for cleaving viral polyproteins during replication. Here, we discovered new functions of the NSP3 and NSP5 proteases of SARS-CoV-2, demonstrating that they could directly cleave proteins involved in the host innate immune response. We identified 3 proteins that were specifically and selectively cleaved by NSP3 or NSP5: IRF-3, and NLRP12 and TAB1, respectively. Direct cleavage of IRF3 by NSP3 could explain the blunted Type-I IFN response seen during SARS-CoV-2 infections while NSP5 mediated cleavage of NLRP12 and TAB1 point to a molecular mechanism for enhanced production of cytokines and inflammatory responThe genome of SARS-CoV-2 encodes two viral proteases (NSP3/papain-like protease and NSP5/3C-like protease) that are responsible for cleaving viral polyproteins during replication. Here, we discovered new functions of the NSP3 and NSP5 proteases of SARS-CoV-2, demonstrating that they could directly cleave proteins involved in the host innate immune response. We identified 3 proteins that were specifically and selectively cleaved by NSP3 or NSP5: IRF-3, and NLRP12 and TAB1, respectively. Direct cleavage of IRF3 by NSP3 could explain the blunted Type-I IFN response seen during SARS-CoV-2 infections while NSP5 mediated cleavage of NLRP12 and TAB1 point to a molecular mechanism for enhanced production of cytokines and inflammatory response observed in COVID-19 patients. We demonstrate that in the mouse NLRP12 protein, one of the recognition site is not cleaved in our in-vitro assay. We pushed this comparative alignment of IRF-3 and NLRP12 homologs and show that the lack or presence of cognate cleavage motifs in IRF-3 and NLRP12 could contribute to the presentation of disease in cats and tigers, for example. Our findings provide an explanatory framework for indepth studies into the pathophysiology of COVID-19.
AB - The genome of SARS-CoV-2 encodes two viral proteases (NSP3/papain-like protease and NSP5/3C-like protease) that are responsible for cleaving viral polyproteins during replication. Here, we discovered new functions of the NSP3 and NSP5 proteases of SARS-CoV-2, demonstrating that they could directly cleave proteins involved in the host innate immune response. We identified 3 proteins that were specifically and selectively cleaved by NSP3 or NSP5: IRF-3, and NLRP12 and TAB1, respectively. Direct cleavage of IRF3 by NSP3 could explain the blunted Type-I IFN response seen during SARS-CoV-2 infections while NSP5 mediated cleavage of NLRP12 and TAB1 point to a molecular mechanism for enhanced production of cytokines and inflammatory responThe genome of SARS-CoV-2 encodes two viral proteases (NSP3/papain-like protease and NSP5/3C-like protease) that are responsible for cleaving viral polyproteins during replication. Here, we discovered new functions of the NSP3 and NSP5 proteases of SARS-CoV-2, demonstrating that they could directly cleave proteins involved in the host innate immune response. We identified 3 proteins that were specifically and selectively cleaved by NSP3 or NSP5: IRF-3, and NLRP12 and TAB1, respectively. Direct cleavage of IRF3 by NSP3 could explain the blunted Type-I IFN response seen during SARS-CoV-2 infections while NSP5 mediated cleavage of NLRP12 and TAB1 point to a molecular mechanism for enhanced production of cytokines and inflammatory response observed in COVID-19 patients. We demonstrate that in the mouse NLRP12 protein, one of the recognition site is not cleaved in our in-vitro assay. We pushed this comparative alignment of IRF-3 and NLRP12 homologs and show that the lack or presence of cognate cleavage motifs in IRF-3 and NLRP12 could contribute to the presentation of disease in cats and tigers, for example. Our findings provide an explanatory framework for indepth studies into the pathophysiology of COVID-19.
KW - IRF3
KW - NLRP12
KW - NSP3 (PLpro)
KW - NSP5 (3CLpro)
KW - SARS-CoV-2
KW - TAB1
KW - innate immunity
KW - protease activity
UR - http://www.scopus.com/inward/record.url?scp=85100289034&partnerID=8YFLogxK
U2 - 10.1080/22221751.2020.1870414
DO - 10.1080/22221751.2020.1870414
M3 - Article
C2 - 33372854
AN - SCOPUS:85100289034
SN - 2222-1751
VL - 10
SP - 178
EP - 195
JO - Emerging Microbes and Infections
JF - Emerging Microbes and Infections
IS - 1
ER -