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SARS-CoV-2 NSP12 Protein Is Not an Interferon-ß Antagonist.
Li, Aixin; Zhao, Kaitao; Zhang, Bei; Hua, Rong; Fang, Yujie; Jiang, Wuhui; Zhang, Jing; Hui, Lixia; Zheng, Yingcheng; Li, Yan; Zhu, Chengliang; Wang, Pei-Hui; Peng, Ke; Xia, Yuchen.
  • Li A; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Zhao K; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Zhang B; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Hua R; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Fang Y; State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, Hubei, China.
  • Jiang W; University of Chinese Academy of Sciences, Beijing, China.
  • Zhang J; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Hui L; Key Laboratory for Experimental Teratology of Ministry of Education and Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong Universitygrid.27255.37, Jinan, China.
  • Zheng Y; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Li Y; State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan Universitygrid.49470.3e, Wuhan, Hubei, China.
  • Zhu C; Department of Pathogen Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
  • Wang PH; Tongji-Rongcheng Center for Biomedicine, Huazhong University of Science and Technology, Wuhan, China.
  • Peng K; Department of Clinical Laboratory, Renmin Hospital of Wuhan Universitygrid.49470.3e, Wuhan, China.
  • Xia Y; Key Laboratory for Experimental Teratology of Ministry of Education and Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong Universitygrid.27255.37, Jinan, China.
J Virol ; 95(17): e0074721, 2021 08 10.
Article in English | MEDLINE | ID: covidwho-1350002
Semantic information from SemMedBD (by NLM)
1. 2019 novel coronavirus CAUSES COVID-19
Subject
2019 novel coronavirus
Predicate
CAUSES
Object
COVID-19
2. interferon-beta AFFECTS luciferin monooxygenase activity
Subject
interferon-beta
Predicate
AFFECTS
Object
luciferin monooxygenase activity
3. Sendai virus infection CAUSES interferon-beta production
Subject
Sendai virus infection
Predicate
CAUSES
Object
interferon-beta production
4. Interferon beta assay USES Luciferases
Subject
Interferon beta assay
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USES
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Luciferases
5. Assay USES Tracer
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Assay
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6. Proteins PART_OF 2019 novel coronavirus
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Proteins
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PART_OF
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2019 novel coronavirus
7. Proteins DISRUPTS Interferon Signaling Process
Subject
Proteins
Predicate
DISRUPTS
Object
Interferon Signaling Process
8. interferon-beta NEG_DISRUPTS interferon-beta production
Subject
interferon-beta
Predicate
NEG_DISRUPTS
Object
interferon-beta production
9. Assay USES Luciferases
Subject
Assay
Predicate
USES
Object
Luciferases
10. 2019 novel coronavirus CAUSES COVID-19
Subject
2019 novel coronavirus
Predicate
CAUSES
Object
COVID-19
11. interferon-beta AFFECTS luciferin monooxygenase activity
Subject
interferon-beta
Predicate
AFFECTS
Object
luciferin monooxygenase activity
12. Sendai virus infection CAUSES interferon-beta production
Subject
Sendai virus infection
Predicate
CAUSES
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interferon-beta production
13. Interferon beta assay USES Luciferases
Subject
Interferon beta assay
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USES
Object
Luciferases
14. Assay USES Tracer
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Assay
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USES
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Tracer
15. Proteins PART_OF 2019 novel coronavirus
Subject
Proteins
Predicate
PART_OF
Object
2019 novel coronavirus
16. Proteins DISRUPTS Interferon Signaling Process
Subject
Proteins
Predicate
DISRUPTS
Object
Interferon Signaling Process
17. interferon-beta NEG_DISRUPTS interferon-beta production
Subject
interferon-beta
Predicate
NEG_DISRUPTS
Object
interferon-beta production
18. Assay USES Luciferases
Subject
Assay
Predicate
USES
Object
Luciferases
ABSTRACT
The coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is bringing an unprecedented health crisis to the world. To date, our understanding of the interaction between SARS-CoV-2 and host innate immunity is still limited. Previous studies reported that SARS-CoV-2 nonstructural protein 12 (NSP12) was able to suppress interferon-ß (IFN-ß) activation in IFN-ß promoter luciferase reporter assays, which provided insights into the pathogenesis of COVID-19. In this study, we demonstrated that IFN-ß promoter-mediated luciferase activity was reduced during coexpression of NSP12. However, we could show NSP12 did not affect IRF3 or NF-κB activation. Moreover, IFN-ß production induced by Sendai virus (SeV) infection or other stimulus was not affected by NSP12 at mRNA or protein level. Additionally, the type I IFN signaling pathway was not affected by NSP12, as demonstrated by the expression of interferon-stimulated genes (ISGs). Further experiments revealed that different experiment systems, including protein tags and plasmid backbones, could affect the readouts of IFN-ß promoter luciferase assays. In conclusion, unlike as previously reported, our study showed SARS-CoV-2 NSP12 protein is not an IFN-ß antagonist. It also rings the alarm on the general usage of luciferase reporter assays in studying SARS-CoV-2. IMPORTANCE Previous studies investigated the interaction between SARS-CoV-2 viral proteins and interferon signaling and proposed that several SARS-CoV-2 viral proteins, including NSP12, could suppress IFN-ß activation. However, most of these results were generated from IFN-ß promoter luciferase reporter assay and have not been validated functionally. In our study, we found that, although NSP12 could suppress IFN-ß promoter luciferase activity, it showed no inhibitory effect on IFN-ß production or its downstream signaling. Further study revealed that contradictory results could be generated from different experiment systems. On one hand, we demonstrated that SARS-CoV-2 NSP12 could not suppress IFN-ß signaling. On the other hand, our study suggests that caution needs to be taken with the interpretation of SARS-CoV-2-related luciferase assays.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Promoter Regions, Genetic / Interferon-beta / Coronavirus RNA-Dependent RNA Polymerase / SARS-CoV-2 Limits: Humans Language: English Journal: J Virol Year: 2021 Document Type: Article Affiliation country: JVI.00747-21

Full text: Available Collection: International databases Database: MEDLINE Main subject: Promoter Regions, Genetic / Interferon-beta / Coronavirus RNA-Dependent RNA Polymerase / SARS-CoV-2 Limits: Humans Language: English Journal: J Virol Year: 2021 Document Type: Article Affiliation country: JVI.00747-21