PDCoV nsp14通过自噬和蛋白酶体途径降解MAVS、MyD88和TRAF3蛋白抑制干扰素通路

doi:10.1016/j.jia.2025.04.015

摘要/Abstract

摘要：

猪δ冠状病毒（PDCoV）是一种新发的猪肠道冠状病毒，可引起哺乳仔猪急性腹泻、呕吐、脱水和死亡。该病毒不仅严重危害养猪业的健康发展，其潜在的跨物种传播风险更值得关注。PDCoV的非结构蛋白14（nsp14）抑制β干扰素（IFN-β）的机制不清。在本研究中，我们发现PDCoV的nsp14蛋白能够通过泛素-蛋白酶体和自噬-溶酶体途径，特异性靶向降解干扰素信号通路中的关键接头蛋白MAVS、MyD88和TRAF3。PDCoV nsp14通过招募E3泛素连接酶MARCH8介导MAVS、MyD88和TRAF3蛋白的泛素化修饰。泛素化后的蛋白被货物受体NDP52识别，并转运至溶酶体降解，有效抑制IFN-β的表达。这一过程揭示了PDCoV nsp14在抑制天然免疫和促进病毒复制中的重要作用。我们还发现：宿主细胞中的MAVS、MyD88和TRAF3蛋白也能够通过激活NDP52介导的选择性自噬途径降解PDCoV的nsp14蛋白，抑制nsp14拮抗天然免疫。

本研究揭示了PDCoV nsp14与宿主蛋白之间的相互博弈机制。一方面，PDCoV nsp14通过选择性自噬途径降解宿主蛋白，抑制宿主的天然免疫，为病毒的复制和增殖创造有利条件；另一方面，宿主细胞通过选择性自噬途径降解PDCoV nsp14蛋白，试图清除病毒并维持自身的免疫防御功能。这种双向蛋白降解机制的发现，不仅阐明了PDCoV免疫逃逸的核心策略，更为理解冠状病毒与宿主的协同进化提供了新视角。本研究为开发靶向病毒免疫逃逸机制的广谱抗冠状病毒药物奠定了理论基础。

Abstract:

Porcine deltacoronavirus (PDCoV) is a newly found pathogen that could potentially cross-species transmit to threat the safety of swine and human. The mechanism of PDCoV nonstructural protein 14 (nsp14) inhibits the expression of IFN-β is unknown. In this study, we showed that PDCoV nsp14 degrades MAVS, MyD88 and TRAF3 protein in host cells by proteasomal and autophagy pathway. PDCoV nsp14 recruites E3 ubiquitin ligase MARCH8 for catalyzing MAVS, MyD88 and TRAF3 protein ubiquitination, and which were recognized and transported to lysosome by the cargo receptor NDP52 for degradation to inhibit the expression of IFN-β. Furthermore, we found that MAVS, MyD88 and TRAF3 also degrade PDCoV nsp14 by selective autophagy. These results reveal the dual function of selective autophagy in PDCoV nsp14 and host proteins, which could promote the ubiquitination of viral particles and host antiviral proteins to degrade both of the proteins for regulating the relationship between virus infection and host innate immunity.

Key words: PDCoV , nsp14 , interferon , autophagosome , proteasome

. PDCoV nsp14通过自噬和蛋白酶体途径降解MAVS、MyD88和TRAF3蛋白抑制干扰素通路[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2025.04.015.

Yiyi Song, Ning Kong, Lanlan Zheng, Yu Zhang, Xueying Zhai, Wenzhen Qin, Xinyu Yang, Xiaoquan Wang, Ao Gao, Wu Tong, Changlong Liu, Hao Zheng, Hai Yu, Wen Zhang, Guangzhi Tong, Tongling Shan. PDCoV nsp14 interferes with the interferon pathway by degrading MAVS, MyD88, and TRAF3 proteins via the autophagy and proteasome pathways[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2025.04.015.

参考文献

Boley P A, Alhamo M A, Lossie G, Yadav K K, Vasquez-Lee M, Saif L J, Kenney S P. 2020. Porcine Deltacoronavirus Infection and Transmission in Poultry, United States(1). Emerging Infectious Disease, 26, 255-265.

Bourgeois C, Majer O, Frohner I E, Lesiak-Markowicz I, Hildering K S, Glaser W, Stockinger S, Decker T, Akira S, Müller M, Kuchler K. 2011. Conventional dendritic cells mount a type I IFN response against Candida spp. requiring novel phagosomal TLR7-mediated IFN-β signaling. Journal of Immunology, 186, 3104-3112.

Chen F, Zhu Y, Wu M, Ku X, Yao L, He Q. 2015. Full-Length Genome Characterization of Chinese Porcine Deltacoronavirus Strain CH/SXD1/2015. Genome Announcements, 3.

Chen J, Fang P, Wang M, Peng Q, Ren J, Wang D, Peng G, Fang L, Xiao S, Ding Z. 2019. Porcine deltacoronavirus nucleocapsid protein antagonizes IFN-β production by impairing dsRNA and PACT binding to RIG-I. Virus Genes, 55, 520-531.

Ding Z, Fang L, Jing H, Zeng S, Wang D, Liu L, Zhang H, Luo R, Chen H, Xiao S. 2014. Porcine epidemic diarrhea virus nucleocapsid protein antagonizes beta interferon production by sequestering the interaction between IRF3 and TBK1. Journal of Virology, 88, 8936-8945.

Dong N, Fang L, Yang H, Liu H, Du T, Fang P, Wang D, Chen H, Xiao S. 2016. Isolation, genomic characterization, and pathogenicity of a Chinese porcine deltacoronavirus strain CHN-HN-2014. Veterinary Microbiology, 196, 98-106.

Dong S, Kong N, Zhang Y, Li Y, Sun D, Qin W, Zhai H, Zhai X, Yang X, Ye C, Ye M, Liu C, Yu L, Zheng H, Tong W, Yu H, Zhang W, Tong G, Shan T. 2022. TARDBP Inhibits Porcine Epidemic Diarrhea Virus Replication through Degrading Viral Nucleocapsid Protein and Activating Type I Interferon Signaling. Journal of Virology, 96, e0007022.

Duan C. 2021. An Updated Review of Porcine Deltacoronavirus in Terms of Prevalence, Pathogenicity, Pathogenesis and Antiviral Strategy. Frontiers in Veterinary Science, 8, 811187.

Duan C, Liu Y, Hao Z, Wang J. 2021. Ergosterol peroxide suppresses porcine deltacoronavirus (PDCoV)-induced autophagy to inhibit virus replication via p38 signaling pathway. Veterinary Microbiology, 257, 109068.

Eckerle L D, Becker M M, Halpin R A, Li K, Venter E, Lu X, Scherbakova S, Graham R L, Baric R S, Stockwell T B, Spiro D J, Denison M R. 2010. Infidelity of SARS-CoV Nsp14-exonuclease mutant virus replication is revealed by complete genome sequencing. PLoS Pathogens, 6, e1000896.

Fang P, Fang L, Xia S, Ren J, Zhang J, Bai D, Zhou Y, Peng G, Zhao S, Xiao S. 2020. Porcine Deltacoronavirus Accessory Protein NS7a Antagonizes IFN-β Production by Competing With TRAF3 and IRF3 for Binding to IKKε. Frontiers in Cellular and Infection Microbiology, 10, 257.

Fitzgerald K A, Kagan J C. 2020. Toll-like Receptors and the Control of Immunity. Cell, 180, 1044-1066.

Fitzgerald K A, McWhirter S M, Faia K L, Rowe D C, Latz E, Golenbock D T, Coyle A J, Liao S M, Maniatis T. 2003. IKKepsilon and TBK1 are essential components of the IRF3 signaling pathway. Nature Immunology, 4, 491-496.

Gribble J, Stevens L J, Agostini M L, Anderson-Daniels J, Chappell J D, Lu X, Pruijssers A J, Routh A L, Denison M R. 2021. The coronavirus proofreading exoribonuclease mediates extensive viral recombination. PLoS Pathogens, 17, e1009226.

He W T, Ji X, He W, Dellicour S, Wang S, Li G, Zhang L, Gilbert M, Zhu H, Xing G, Veit M, Huang Z, Han G Z, Huang Y, Suchard M A, Baele G, Lemey P, Su S. 2020. Genomic Epidemiology, Evolution, and Transmission Dynamics of Porcine Deltacoronavirus. Molecular Biology and Evolution, 37, 2641-2654.

Hu H, Jung K, Vlasova A N, Chepngeno J, Lu Z, Wang Q, Saif L J. 2015. Isolation and characterization of porcine deltacoronavirus from pigs with diarrhea in the United States. Journal of Clinical Microbiology, 53, 1537-1548.

Ivashkiv L B, Donlin L T. 2014. Regulation of type I interferon responses. Nature Reviews Immunology, 14, 36-49.

Jaru-Ampornpan P, Jengarn J, Wanitchang A, Jongkaewwattana A. 2017. Porcine Epidemic Diarrhea Virus 3C-Like Protease-Mediated Nucleocapsid Processing: Possible Link to Viral Cell Culture Adaptability. Journal of Virology, 91.

Jiao Y, Kong N, Wang H, Sun D, Dong S, Chen X, Zheng H, Tong W, Yu H, Yu L, Huang Y, Wang H, Sui B, Zhao L, Liao Y, Zhang W, Tong G, Shan T. 2021. PABPC4 Broadly Inhibits Coronavirus Replication by Degrading Nucleocapsid Protein through Selective Autophagy. Microbiology Spectrum, 9, e0090821.

Kawai T, Takahashi K, Sato S, Coban C, Kumar H, Kato H, Ishii K J, Takeuchi O, Akira S. 2005. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nature Immunology, 6, 981-988.

Kong F, Wang Q, Kenney S P, Jung K, Vlasova A N, Saif L J. 2022. Porcine Deltacoronaviruses: Origin, Evolution, Cross-Species Transmission and Zoonotic Potential. Pathogens, 11.

Kong N, Shan T, Wang H, Jiao Y, Zuo Y, Li L, Tong W, Yu L, Jiang Y, Zhou Y, Li G, Gao F, Yu H, Zheng H, Tong G. 2020. BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy. Autophagy, 16, 1737-1752.

Kraft C, Peter M, Hofmann K. 2010. Selective autophagy: ubiquitin-mediated recognition and beyond. Nature Cell Biology, 12, 836-841.

Lednicky J A, Tagliamonte M S, White S K, Elbadry M A, Alam M M, Stephenson C J, Bonny T S, Loeb J C, Telisma T, Chavannes S, Ostrov D A, Mavian C, Beau De Rochars V M, Salemi M, Morris J G, Jr. 2021. Independent infections of porcine deltacoronavirus among Haitian children. Nature, 600, 133-137.

Li G, Chen Q, Harmon K M, Yoon K J, Schwartz K J, Hoogland M J, Gauger P C, Main R G, Zhang J. 2014. Full-Length Genome Sequence of Porcine Deltacoronavirus Strain USA/IA/2014/8734. Genome Announcements, 2.

Liu X, Fang P, Fang L, Hong Y, Zhu X, Wang D, Peng G, Xiao S. 2019. Porcine deltacoronavirus nsp15 antagonizes interferon-β production independently of its endoribonuclease activity. Molecular Immunology, 114, 100-107.

Liu Y, Liang Q Z, Lu W, Yang Y L, Chen R, Huang Y W, Wang B. 2021. A Comparative Analysis of Coronavirus Nucleocapsid (N) Proteins Reveals the SADS-CoV N Protein Antagonizes IFN-β Production by Inducing Ubiquitination of RIG-I. Frontiers in Immunology, 12, 688758.

Ma Y, Wu L, Shaw N, Gao Y, Wang J, Sun Y, Lou Z, Yan L, Zhang R, Rao Z. 2015. Structural basis and functional analysis of the SARS coronavirus nsp14-nsp10 complex. Proceedings of the National Academy of Sciences of the United States of America, 112, 9436-9441.

Meylan E, Curran J, Hofmann K, Moradpour D, Binder M, Bartenschlager R, Tschopp J. 2005. Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature, 437, 1167-1172.

Meylan E, Tschopp J, Karin M. 2006. Intracellular pattern recognition receptors in the host response. Nature, 442, 39-44.

Miller K, McGrath M E, Hu Z, Ariannejad S, Weston S, Frieman M, Jackson W T. 2020. Coronavirus interactions with the cellular autophagy machinery. Autophagy, 16, 2131-2139.

Mizushima N, Komatsu M. 2011. Autophagy: renovation of cells and tissues. Cell, 147, 728-741.

Mizushima N, Yoshimori T, Levine B. 2010. Methods in mammalian autophagy research. Cell, 140, 313-326.

Qin W, Kong N, Zhang Y, Dong S, Zhai H, Zhai X, Yang X, Ye C, Ye M, Liu C, Yu L, Zheng H, Yu H, Zhang W, Tong G, Lan D, Tong W, Shan T. 2022. Nuclear ribonucleoprotein RALY targets virus nucleocapsid protein and induces autophagy to restrict porcine epidemic diarrhea virus replication. Journal of Biological Chemistry, 298, 102190.

Qin W, Kong N, Zhang Y, Wang C, Dong S, Zhai H, Zhai X, Yang X, Ye C, Ye M, Tong W, Liu C, Yu L, Zheng H, Yu H, Zhang W, Lan D, Tong G, Shan T. 2023. PTBP1 suppresses porcine epidemic diarrhea virus replication via inducing protein degradation and IFN production. Journal of Biological Chemistry, 299, 104987.

Saramago M, Bárria C, Costa V G, Souza C S, Viegas S C, Domingues S, Lousa D, Soares C M, Arraiano C M, Matos R G. 2021. New targets for drug design: importance of nsp14/nsp10 complex formation for the 3'-5' exoribonucleolytic activity on SARS-CoV-2. Febs Journal, 288, 5130-5147.

Seth R B, Sun L, Ea C K, Chen Z J. 2005. Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3. Cell, 122, 669-682.

Shi P, Su Y, Li R, Liang Z, Dong S, Huang J. 2019. PEDV nsp16 negatively regulates innate immunity to promote viral proliferation. Virus Research, 265, 57-66.

Stefan K L, Kim M V, Iwasaki A, Kasper D L. 2020. Commensal Microbiota Modulation of Natural Resistance to Virus Infection. Cell, 183, 1312-1324.e1310.

Stetson D B, Medzhitov R. 2006. Type I interferons in host defense. Immunity, 25, 373-381.

Sun L, Liu S, Chen Z J. 2010. SnapShot: pathways of antiviral innate immunity. Cell, 140, 436-436.e432.

Tang P, Cui E, Song Y, Yan R, Wang J. 2021. Porcine deltacoronavirus and its prevalence in China: a review of epidemiology, evolution, and vaccine development. Archives of Virology, 166, 2975-2988.

Tanji T, Ip Y T. 2005. Regulators of the Toll and Imd pathways in the Drosophila innate immune response. Trends in Immunology, 26, 193-198.

Totura A L, Whitmore A, Agnihothram S, Schäfer A, Katze M G, Heise M T, Baric R S. 2015. Toll-Like Receptor 3 Signaling via TRIF Contributes to a Protective Innate Immune Response to Severe Acute Respiratory Syndrome Coronavirus Infection. mBio, 6, e00638-00615.

Wang D, Fang L, Shi Y, Zhang H, Gao L, Peng G, Chen H, Li K, Xiao S. 2016. Porcine Epidemic Diarrhea Virus 3C-Like Protease Regulates Its Interferon Antagonism by Cleaving NEMO. Journal of Virology, 90, 2090-2101.

Wang L, Byrum B, Zhang Y. 2014. Detection and genetic characterization of deltacoronavirus in pigs, Ohio, USA, 2014. Emerging Infectious Disease, 20, 1227-1230.

Woo P C, Lau S K, Lam C S, Lau C C, Tsang A K, Lau J H, Bai R, Teng J L, Tsang C C, Wang M, Zheng B J, Chan K H, Yuen K Y. 2012. Discovery of seven novel Mammalian and avian coronaviruses in the genus deltacoronavirus supports bat coronaviruses as the gene source of alphacoronavirus and betacoronavirus and avian coronaviruses as the gene source of gammacoronavirus and deltacoronavirus. Journal of Virology, 86, 3995-4008.

Yang Y L, Meng F, Qin P, Herrler G, Huang Y W, Tang Y D. 2020. Trypsin promotes porcine deltacoronavirus mediating cell-to-cell fusion in a cell type-dependent manner. Emerging Microbes & Infections, 9, 457-468.

Yin L, Chen J, Li L, Guo S, Xue M, Zhang J, Liu X, Feng L, Liu P. 2020. Aminopeptidase N Expression, Not Interferon Responses, Determines the Intestinal Segmental Tropism of Porcine Deltacoronavirus. Journal of Virology, 94.

Zhai X, Kong N, Zhang Y, Song Y, Qin W, Yang X, Ye C, Ye M, Tong W, Liu C, Zheng H, Yu H, Zhang W, Yang X, Zhang G, Tong G, Shan T. 2023. N protein of PEDV plays chess game with host proteins by selective autophagy. Autophagy, 19, 2338-2352.

Zhang Q, Yoo D. 2016. Immune evasion of porcine enteric coronaviruses and viral modulation of antiviral innate signaling. Virus Research, 226, 128-141.

Zhao D, Gao X, Zhou P, Zhang L, Zhang Y, Wang Y, Liu X. 2020. Evaluation of the immune response in conventionally weaned pigs infected with porcine deltacoronavirus. Archives of Virology, 165, 1653-1658.

Zhou X, Zhou L, Ge X, Guo X, Han J, Zhang Y, Yang H. 2020. Quantitative Proteomic Analysis of Porcine Intestinal Epithelial Cells Infected with Porcine Deltacoronavirus Using iTRAQ-Coupled LC-MS/MS. Journal of Proteome Research, 19, 4470-4485.

Zhu X, Wang D, Zhou J, Pan T, Chen J, Yang Y, Lv M, Ye X, Peng G, Fang L, Xiao S. 2017. Porcine Deltacoronavirus nsp5 Antagonizes Type I Interferon Signaling by Cleaving STAT2. Journal of Virology, 91.