Abstract
A deficient interferon (IFN) response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has been implicated as a determinant of severe coronavirus disease 2019 (COVID-19). To identify the molecular effectors that govern IFN control of SARS-CoV-2 infection, we conducted a large-scale gain-of-function analysis that evaluated the impact of human IFN-stimulated genes (ISGs) on viral replication. A limited subset of ISGs were found to control viral infection, including endosomal factors inhibiting viral entry, RNA binding proteins suppressing viral RNA synthesis, and a highly enriched cluster of endoplasmic reticulum (ER)/Golgi-resident ISGs inhibiting viral assembly/egress. These included broad-acting antiviral ISGs and eight ISGs that specifically inhibited SARS-CoV-2 and SARS-CoV-1 replication. Among the broad-acting ISGs was BST2/tetherin, which impeded viral release and is antagonized by SARS-CoV-2 Orf7a protein. Overall, these data illuminate a set of ISGs that underlie innate immune control of SARS-CoV-2/SARS-CoV-1 infection, which will facilitate the understanding of host determinants that impact disease severity and offer potential therapeutic strategies for COVID-19.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2656-2668.e8 |
| Journal | Molecular cell |
| Volume | 81 |
| Issue number | 12 |
| DOIs | |
| State | Published - Jun 17 2021 |
Funding
We would like to thank Ralph Baric for providing the dOrf7a SARS-CoV-2, Thomas Rogers for the HeLa-ACE2 cells, Kwok-Yung Yuen for the rabbit-anti-SARS-CoV-2 N antibody, and Stefan Pohlmann for the mammalian expression vector encoding SARS-CoV-2 S and pCG1-CoV2-S-HA. We thank Marisol Chacon for administrative support and Sylvie Blondelle and Larry Adelman for biosafety support. We would also like to thank the Viral Vector Core Facility at SBP for cDNA normalization and lentivirus production. This work was supported by the following grants to the SBP Medical Discovery Institute and the Icahn School of Medicine at Mount Sinai: DOD W81XWH-20-1-0270, DHIPC U19 AI118610, and Fluomics/NOSI U19 AI135972. This work was also supported by generous philanthropic donations from Dinah Ruch and Susan & James Blair, the JPB Foundation, the Open Philanthropy Project (research grant 2020-215611 (5384)), and anonymous donors. Additional support has been provided by DARPA (grant HR0011-19-2-0020) and CRIP (Center for Research on Influenza Pathogenesis), a NIAID-funded Center of Excellence for Influenza Research and Surveillance (CEIRS; contract HHSN272201400008C). This work was additionally supported by the following grants to Northwestern University Feinberg School of Medicine: a CTSA supplement to NCATS (UL1 TR002389), a CTSA supplement to NUCATS with the generous support of the Dixon family (UL1 TR001422), and a Cancer Center supplement (P30 CA060553). Additional support was provided by NIH grant R37AI081668 to J.G. at UC San Diego and a generous grant from the James B. Pendleton Charitable Trust. The funding sources had no role in the study design, data collection, analysis, interpretation, or writing of the report. The content of this study is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. L.M.-S. M.K.L. C.A.S. J.G. and S.K.C. conceived and designed the experiments. L.M.-S. M.K.L. X.Y. C.A.S. A.P.G. P.D.J. C.N. Y.P. M.E.B. S.W. H.M.C. and A.L.O. conducted and/or analyzed the experiments. L.M.-S. L.P. and A.M.O. conducted data analysis and representation. A.R.-F. M.U. M.W.C. and C.B. performed and/or analyzed the RNA-seq experiments. L.M.-S. D.P. C.C. S.L. S.B.R. F.Z. and T.I. generated the network model. C.W. and Y.X. generated the in silico docking model. L.M. and J.F.H. generated essential reagents. L.M.-S. and S.K.C. wrote the manuscript with contributions from all authors. L.P. C.B. A.G.-S. and S.K.C. acquired funding. The authors declare no competing interests. We would like to thank Ralph Baric for providing the dOrf7a SARS-CoV-2, Thomas Rogers for the HeLa-ACE2 cells, Kwok-Yung Yuen for the rabbit-anti-SARS-CoV-2 N antibody, and Stefan Pohlmann for the mammalian expression vector encoding SARS-CoV-2 S and pCG1-CoV2-S-HA. We thank Marisol Chacon for administrative support and Sylvie Blondelle and Larry Adelman for biosafety support. We would also like to thank the Viral Vector Core Facility at SBP for cDNA normalization and lentivirus production. This work was supported by the following grants to the SBP Medical Discovery Institute and the Icahn School of Medicine at Mount Sinai: DOD W81XWH-20-1-0270 , DHIPC U19 AI118610 , and Fluomics/NOSI U19 AI135972 . This work was also supported by generous philanthropic donations from Dinah Ruch and Susan & James Blair , the JPB Foundation , the Open Philanthropy Project (research grant 2020-215611 (5384) ), and anonymous donors. Additional support has been provided by DARPA (grant HR0011-19-2-0020 ) and CRIP ( Center for Research on Influenza Pathogenesis ), a NIAID -funded Center of Excellence for Influenza Research and Surveillance (CEIRS; contract HHSN272201400008C ). This work was additionally supported by the following grants to Northwestern University Feinberg School of Medicine: a CTSA supplement to NCATS ( UL1 TR002389 ), a CTSA supplement to NUCATS with the generous support of the Dixon family ( UL1 TR001422 ), and a Cancer Center supplement ( P30 CA060553 ). Additional support was provided by NIH grant R37AI081668 to J.G. at UC San Diego and a generous grant from the James B. Pendleton Charitable Trust . The funding sources had no role in the study design, data collection, analysis, interpretation, or writing of the report. The content of this study is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
Keywords
- BST2
- ISG
- Orf7a
- SARS-CoV-2
- innate immunity
- interferon
- viral evasion
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology
