SARS-COV-2 protein NSP9 promotes cytokine production by targeting TBK1

Yihua Zhang; Bowen Xin; Yinan Liu; Wenyi Jiang; Wendong Han; Jian Deng; Peihui Wang; Xiaowu Hong; Dapeng Yan

doi:10.3389/fimmu.2023.1211816

SARS-COV-2 protein NSP9 promotes cytokine production by targeting TBK1

Front Immunol. 2023 Oct 2:14:1211816. doi: 10.3389/fimmu.2023.1211816. eCollection 2023.

Authors

Yihua Zhang¹, Bowen Xin¹, Yinan Liu¹, Wenyi Jiang¹, Wendong Han², Jian Deng³, Peihui Wang³, Xiaowu Hong¹, Dapeng Yan¹

Affiliations

¹ Department of Immunology, School of Basic Medical Sciences, Shanghai Institute of Infectious Disease and Biosecurity & Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
² Biosafety Level 3 Laboratory, Fudan University, Shanghai, China.
³ Key Laboratory for Experimental Teratology of Ministry of Education and Advanced Medical Research Institute, Cheeloo College of Medicine, Shandong University, Jinan, China.

Abstract

SARS-COV-2 infection-induced excessive or uncontrolled cytokine storm may cause injury of host tissue or even death. However, the mechanism by which SARS-COV-2 causes the cytokine storm is unknown. Here, we demonstrated that SARS-COV-2 protein NSP9 promoted cytokine production by interacting with and activating TANK-binding kinase-1 (TBK1). With an rVSV-NSP9 virus infection model, we discovered that an NSP9-induced cytokine storm exacerbated tissue damage and death in mice. Mechanistically, NSP9 promoted the K63-linked ubiquitination and phosphorylation of TBK1, which induced the activation and translocation of IRF3, thereby increasing downstream cytokine production. Moreover, the E3 ubiquitin ligase Midline 1 (MID1) facilitated the K48-linked ubiquitination and degradation of NSP9, whereas virus infection inhibited the interaction between MID1 and NSP9, thereby inhibiting NSP9 degradation. Additionally, we identified Lys59 of NSP9 as a critical ubiquitin site involved in the degradation. These findings elucidate a previously unknown mechanism by which a SARS-COV-2 protein promotes cytokine storm and identifies a novel target for COVID-19 treatment.

Keywords: SARS-CoV-2; TBK1; antiviral immunity; cytokine storm; type I interferon.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
COVID-19 Drug Treatment
COVID-19* / complications
COVID-19* / genetics
COVID-19* / immunology
Cytokine Release Syndrome* / etiology
Cytokine Release Syndrome* / genetics
Cytokine Release Syndrome* / immunology
Cytokines
Disease Models, Animal
Immunity, Innate
Mice
Protein Serine-Threonine Kinases* / genetics
Protein Serine-Threonine Kinases* / metabolism
SARS-CoV-2* / genetics
SARS-CoV-2* / immunology
SARS-CoV-2* / metabolism
Signal Transduction
Ubiquitin-Protein Ligases / metabolism

Substances

Cytokines
NSP9 protein, SARS-CoV-2
Protein Serine-Threonine Kinases
Tbk1 protein, mouse
Ubiquitin-Protein Ligases

Grants and funding

This work was supported by grants from the Natural Science Foundation of Shanghai (21ZR1481400), the National Natural Science Foundation of China (32270973 and 31972900), Shanghai Pilot Program for Basic Research (21TQ017), the Program of Shanghai Academic Research Leader (21XD1402900) and the National Youth Talent Support Program (Ten Thousand Talent Program).