E3 ligase Nedd4l promotes antiviral innate immunity by catalyzing K29-linked cysteine ubiquitination of TRAF3

Peng Gao; Xianwei Ma; Ming Yuan; Yulan Yi; Guoke Liu; Mingyue Wen; Wei Jiang; Ruihua Ji; Lingxi Zhu; Zhen Tang; Qingzhuo Yu; Jing Xu; Rui Yang; Sheng Xia; Mingjin Yang; Jianping Pan; Hongbin Yuan; Huazhang An

doi:10.1038/s41467-021-21456-1

E3 ligase Nedd4l promotes antiviral innate immunity by catalyzing K29-linked cysteine ubiquitination of TRAF3

Nat Commun. 2021 Feb 19;12(1):1194. doi: 10.1038/s41467-021-21456-1.

Authors

Peng Gao^#¹, Xianwei Ma^#², Ming Yuan^#³, Yulan Yi^#¹, Guoke Liu¹, Mingyue Wen³, Wei Jiang¹, Ruihua Ji⁴, Lingxi Zhu¹, Zhen Tang¹, Qingzhuo Yu¹, Jing Xu¹, Rui Yang⁴, Sheng Xia⁵, Mingjin Yang³, Jianping Pan⁶, Hongbin Yuan⁷, Huazhang An⁸

Affiliations

¹ Clinical Cancer Institute, Center for Translational Medicine, Second Military Medical University, Shanghai, 200433, China.
² Scientific Research Center, Shanghai Public Health Clinical Center, Fudan University, Shanghai, 201508, China.
³ Immunology Department & National Key Laboratory of Medical Immunology, Second Military Medical University, Shanghai, 200433, China.
⁴ Department of Anesthesiology, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China.
⁵ Department of Immunology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu, 212013, China.
⁶ Department of Clinical Medicine, Zhejiang University City College School of Medicine, Hangzhou, 310015, China.
⁷ Department of Anesthesiology, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China. [email protected].
⁸ Clinical Cancer Institute, Center for Translational Medicine, Second Military Medical University, Shanghai, 200433, China. [email protected].

^# Contributed equally.

Abstract

Ubiquitination is one of the most prevalent protein posttranslational modifications. Here, we show that E3 ligase Nedd4l positively regulates antiviral immunity by catalyzing K29-linked cysteine ubiquitination of TRAF3. Deficiency of Nedd4l significantly impairs type I interferon and proinflammatory cytokine production induced by virus infection both in vitro and in vivo. Nedd4l deficiency inhibits virus-induced ubiquitination of TRAF3, the binding between TRAF3 and TBK1, and subsequent phosphorylation of TBK1 and IRF3. Nedd4l directly interacts with TRAF3 and catalyzes K29-linked ubiquitination of Cys56 and Cys124, two cysteines that constitute zinc fingers, resulting in enhanced association between TRAF3 and E3 ligases, cIAP1/2 and HECTD3, and also increased K48/K63-linked ubiquitination of TRAF3. Mutation of Cys56 and Cys124 diminishes Nedd4l-catalyzed K29-linked ubiquitination, but enhances association between TRAF3 and the E3 ligases, supporting Nedd4l promotes type I interferon production in response to virus by catalyzing ubiquitination of the cysteines in TRAF3.

Publication types

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

MeSH terms

Animals
Antiviral Agents / pharmacology*
Baculoviral IAP Repeat-Containing 3 Protein / metabolism
Catalysis
Cysteine / metabolism
Female
HEK293 Cells
Humans
Immunity, Innate / drug effects*
Inhibitor of Apoptosis Proteins / metabolism
Interferon Regulatory Factor-3 / metabolism
Interferon Type I / metabolism
Mice
Mice, Inbred C57BL
Mice, Knockout
Nedd4 Ubiquitin Protein Ligases / deficiency
Nedd4 Ubiquitin Protein Ligases / genetics*
Nedd4 Ubiquitin Protein Ligases / metabolism*
Protein Processing, Post-Translational
Protein Serine-Threonine Kinases
TNF Receptor-Associated Factor 3 / genetics
TNF Receptor-Associated Factor 3 / metabolism*
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism*
Ubiquitination*

Substances

Antiviral Agents
Inhibitor of Apoptosis Proteins
Interferon Regulatory Factor-3
Interferon Type I
Irf3 protein, mouse
TNF Receptor-Associated Factor 3
HECTd3 protein, mouse
Nedd4 Ubiquitin Protein Ligases
Nedd4l protein, mouse
Baculoviral IAP Repeat-Containing 3 Protein
Ubiquitin-Protein Ligases
Tbk1 protein, mouse
Protein Serine-Threonine Kinases
Cysteine