ZNF219 inhibits porcine epidemic diarrhea virus by degrading viral S2 protein

Wenli Zhao; Ning Kong; Daoliang Lan; Yu Zhang; Hailong Liu; Chunyun Liu; Wenzhen Qin; Xinyu Yang; Tian Liu; Wu Tong; Hai Yu; Hao Zheng; Guangzhi Tong; Shengsong Xie; Guangxu Xing; Tongling Shan

doi:10.1016/j.virs.2026.03.007

April 2026

. doi: 10.1016/j.virs.2026.03.007

Citation: Wenli Zhao, Ning Kong, Daoliang Lan, Yu Zhang, Hailong Liu, Chunyun Liu, Wenzhen Qin, Xinyu Yang, Tian Liu, Wu Tong, Hai Yu, Hao Zheng, Guangzhi Tong, Shengsong Xie, Guangxu Xing, Tongling Shan. ZNF219 inhibits porcine epidemic diarrhea virus by degrading viral S2 protein .VIROLOGICA SINICA, 2026, 41(2) : 303-313. http://dx.doi.org/10.1016/j.virs.2026.03.007

ZNF219通过降解猪流行性腹泻病毒S2蛋白抑制猪流行性腹泻病毒

赵文丽 ^a,b ,
孔宁 ^a,c ,
兰道亮 ^b ,
张宇 ^e ,
刘海龙 ^d ,
刘春雲 ^a,b ,
秦文珍 ^a ,
杨心雨 ^a ,
刘甜 ^a ,
童武 ^a ,
于海 ^a ,
郑浩 ^a ,
童光志 ^a ,
谢胜松 ^d,, ,
邢广旭 ^f,, ,
单同领 ^a,c,,

a 中国农业科学院上海兽医研究所, 上海;
b 西南民族大学动物兽医学院, 成都;
c 扬州大学江苏省重大动物传染病与人畜共患病防控协同创新中心, 扬州;
d 华中农业大学农用动物遗传育种与繁殖教育部重点实验室/农业农村部猪遗传育种重点实验室, 武汉;
e 上海交通大学医学院口腔医学院, 上海市第九人民医院口腔预防科, 上海;
f 河南省农业科学院动物免疫学重点实验室, 郑州

通讯作者： 谢胜松, ssxie@mail.hzau.edu.cn; 邢广旭, xingguangxu@163.com; 单同领, shantongling@shvri.ac.cn
收稿日期： 2025-05-16
录用日期： 2026-03-18

摘要

猪流行性腹泻病毒（PEDV）自发现以来，已对全球农业经济造成重大影响。现有的商业化冠状病毒疫苗无法有效防控新发流行毒株。因此，阐明病毒与抗病毒宿主因子的相关性是必要的。研究发现，病毒非结构蛋白12（nsp12）在PEDV感染后上调宿主蛋白ZNF219。此外，ZNF219通过选择性自噬降解PEDV S2蛋白抑制PEDV的复制。ZNF219招募E3泛素化连接酶TRAF6，使PEDV S2蛋白泛素化。泛素化的PEDV S2蛋白被识别后通过货物受体p62被递送到自噬酶体中进行自噬降解，从而抑制PEDV的增殖。综上所述，宿主细胞通过识别病毒nsp12蛋白感知PEDV感染后，上调ZNF219在细胞内的表达，ZNF219通过激活自噬降解病毒S2蛋白，从而抑制病毒复制。本研究揭示了一种涉及ZNF219的新型抗病毒机制，为预防和治疗PEDV提供了新的靶点。
- ZNF219
- , PEDV
- , TRAF6
- , P62
- , PEDV S2蛋白
- , 选择性自噬

ZNF219 inhibits porcine epidemic diarrhea virus by degrading viral S2 protein

Wenli Zhao ^a,b ,
Ning Kong ^a,c ,
Daoliang Lan ^b ,
Yu Zhang ^e ,
Hailong Liu ^d ,
Chunyun Liu ^a,b ,
Wenzhen Qin ^a ,
Xinyu Yang ^a ,
Tian Liu ^a ,
Wu Tong ^a ,
Hai Yu ^a ,
Hao Zheng ^a ,
Guangzhi Tong ^a ,
Shengsong Xie ^d,, ,
Guangxu Xing ^f,, ,
Tongling Shan ^a,c,,

a. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China;
b. College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China;
c. Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou University, Yangzhou 225009, China;
d. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, China;
e. Department of Preventive Dentistry, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China;
f. Institute for Animal Health, Henan Academy of Agricultural Sciences, Key Laboratory of Animal Immunology of the Ministry of Agriculture, Zhengzhou 450002, China

Corresponding author: Shengsong Xie, ssxie@mail.hzau.edu.cn Guangxu Xing, xingguangxu@163.com Tongling Shan, shantongling@shvri.ac.cn
Received Date: 16 May 2025
Accepted Date: 18 March 2026

Abstract

Since its discovery, porcine epidemic diarrhea virus (PEDV) has significantly affected the agricultural economy worldwide. The available commercialized coronavirus vaccines cannot adequately control emerging strains. Therefore, investigating the correlation between viruses and antiviral host factors is necessary. In this study, we showed that zinc finger protein 219 (ZNF219) was upregulated by viral nonstructural protein 12 (nsp12) upon PEDV challenge. Moreover, ZNF219 inhibited the replication of PEDV through selective autophagic degradation of the PEDV S2 protein. ZNF219 recruited TRAF6, the ubiquitin E3 ligase, to ubiquitinate the PEDV S2 protein. After recognition, the ubiquitinated PEDV S2 protein was delivered to autolysosomes via the cargo receptor p62 for degradation by autophagy, thus inhibiting the proliferation of PEDV. To summarize, after sensing PEDV infection by recognizing the viral nsp12 protein, host cells upregulated the intracellular expression of ZNF219, which degraded the viral S2 protein by activating autophagy, thus suppressing viral replication. Our study revealed a novel antiviral mechanism involving ZNF219 and provided a novel target for preventing and treating PEDV.
- Zinc finger protein 219 (ZNF219)
- , Porcine epidemic diarrhea virus (PEDV)
- , TRAF6
- , P62
- , PEDV S2 protein
- , Selective autophagy

References
1. Brayer, K.J., Kulshreshtha, S.,Segal, D.J., 2008. The protein-binding potential of C2H2 zinc finger domains. Cell Biochem. Biophys., 51, 9-19.
2. Brayer, K.J.,Segal, D.J., 2008. Keep your fingers off my DNA: protein-Protein interactions mediated by C2H2 zinc finger domains. Cell Biochem. Biophys., 50, 111-131.
3. Brian, D.A.,Baric, R.S., 2005. Coronavirus genome structure and replication. Coronavirus Replicat. Reverse Genet., 287, 1-30.
4. Chen, Q., Li, G.W., Stasko, J., Thomas, J.T., Stensland, W.R., Pillatzki, A.E., Gauger, P.C., Schwartz, K.J., Madson, D., Yoon, K.J., Stevenson, G.W., Burrough, E.R., Harmon, K.M., Main, R.G.,Zhang, J.Q., 2014. Isolation and characterization of porcine epidemic diarrhea viruses associated with the 2013 disease outbreak among swine in the United States. J. Clin. Microbiol., 52, 234-243.
5. Choi, Y., Bowman, J.W.,Jung, J.U., 2018. Autophagy during viral infection - a double-edged sword. Nat. Rev. Microbiol., 16, 340-353.
6. Crawford, K., Lager, K.M., Kulshreshtha, V., Miller, L.C.,Faaberg, K.S., 2016. Status of vaccines for porcine epidemic diarrhea virus in the United States and Canada. Virus Res., 226, 108-116.
7. Cruz, D.J.M., Kim, C.J.,Shin, H.J., 2008. The GPRLQPY motif located at the carboxy-terminal of the spike protein induces antibodies that neutralize. Virus Res., 132, 192-196.
8. Duarte, M., Tobler, K., Bridgen, A., Rasschaert, D., Ackermann, M.,Laude, H., 1994. Sequence analysis of the porcine epidemic diarrhea virus genome between the nucleocapsid and spike protein genes reveals a polymorphic ORF. Virology, 198, 466-476.
9. El Abdellaoui-Soussi, F., Yunes-Leites, P.S., Lopez-Maderuelo, D., Garcia-Marques, F., Vazquez, J., Redondo, J.M.,Gomez-Del Arco, P., 2022. Interplay between the Chd4/NuRD complex and the transcription factor Znf219 controls cardiac cell identity. Int. J. Mol. Sci., 23.
10. Gao, Q., Zheng, Z., Wang, H., Yi, S., Zhang, G.,Gong, L., 2021. The new porcine epidemic diarrhea virus outbreak may mean that existing commercial vaccines are not enough to fully protect against the epidemic strains. Front. Vet. Sci., 8, 697839.
11. Glick, D., Barth, S.,Macleod, K.F., 2010. Autophagy: cellular and molecular mechanisms. J. Pathol., 221, 3-12.
12. Graham, R.L.,Baric, R.S., 2010. Recombination, reservoirs, and the modular spike: mechanisms of coronavirus cross-species transmission. J. Virol., 84, 3134-3146.
13. Gubas, A.,Dikic, I., 2022. A guide to horizontal ellipsis the regulation of selective autophagy receptors. FEBS J., 289, 75-89.
14. Hou, P., Wang, X., Wang, H., Wang, T., Yu, Z., Xu, C., Zhao, Y., Wang, W., Zhao, Y., Chu, F., Chang, H., Zhu, H., Lu, J., Zhang, F., Liang, X., Li, X., Wang, S., Gao, Y.,He, H., 2023. The ORF7a protein of SARS-CoV-2 initiates autophagy and limits autophagosome-lysosome fusion via degradation of SNAP29 to promote virus replication. Autophagy, 19, 551-569.
15. Jin, S., Tian, S., Luo, M., Xie, W., Liu, T., Duan, T., Wu, Y.,Cui, J., 2017. Tetherin suppresses type I interferon signaling by targeting MAVS for NDP52-Mediated selective autophagic degradation in human cells. Mol. Cell, 68, 308-322.e304.
16. Johansen, T.,Lamark, T., 2020. Selective autophagy: ATG8 family proteins, LIR motifs and cargo receptors. J. Mol. Biol., 432, 80-103.
17. Kong, N., Shan, T.L., Wang, H., Jiao, Y.J., Zuo, Y.W., Li, L.W., Tong, W., Yu, L.X., Jiang, Y.F., Zhou, Y.J., Li, G.X., Gao, F., Yu, H., Zheng, H.,Tong, G.Z., 2020. BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy. Autophagy, 16, 1737-1752.
18. Kong, N., Wu, Y., Meng, Q., Wang, Z., Zuo, Y., Pan, X., Tong, W., Zheng, H., Li, G., Yang, S., Yu, H., Zhou, E.M., Shan, T.,Tong, G., 2016. Suppression of virulent porcine epidemic diarrhea virus proliferation by the PI3K/Akt/GSK-3α/β pathway. PLoS One, 11, e0161508.
19. Lan, S.H., Wu, S.Y., Zuchini, R., Lin, X.Z., Su, I.J., Tsai, T.F., Lin, Y.J., Wu, C.T.,Liu, H.S., 2014. Autophagy suppresses tumorigenesis of hepatitis B virus-associated hepatocellular carcinoma through degradation of MicroRNA-224. Hepatology, 59, 505-517.
20. Li, F., 2012. Evidence for a common evolutionary origin of coronavirus spike protein receptor-binding subunits. J. Virol., 86, 2856-2858.
21. Li, F., 2016. Structure, function, and evolution of coronavirus spike proteins. Annu. Rev. Virol., 3, 237-261.
22. Li, W.T., Li, H., Liu, Y.B., Pan, Y.F., Deng, F., Song, Y.H., Tang, X.B.,He, Q.G., 2012. New variants of porcine epidemic diarrhea virus, China, 2011. Emerg. Infect. Dis., 18, 1350-1353.
23. Li, W.T., Van Kuppeveld, F.J.M., He, Q.G., Rottier, P.J.M.,Bosch, B.J., 2016. Cellular entry of the porcine epidemic diarrhea virus. Virus Res., 226, 117-127.
24. Meier, T., Timm, M., Montani, M.,Wilkens, L., 2021. Gene networks and transcriptional regulators associated with liver cancer development and progression. BMC Med. Genom., 14.
25. Mizushima, N.,Komatsu, M., 2011. Autophagy: renovation of cells and tissues. Cell, 147, 728-741.
26. Mizushima, N., Yoshimori, T.,Levine, B., 2010. Methods in mammalian autophagy research. Cell, 140, 313-326.
27. Orvedahl, A., Macpherson, S., Sumpter, R., Talloczy, Z., Zou, Z.J.,Levine, B., 2010. Autophagy protects against sindbis virus infection of the central nervous system. Cell Host Microbe, 7, 115-127.
28. Pan, X., Kong, N., Shan, T., Zheng, H., Tong, W., Yang, S., Li, G., Zhou, E.,Tong, G., 2015. Monoclonal antibody to N protein of porcine epidemic diarrhea virus. Monoclon. Antibodies Immunodiagn. Immunother., 34, 51-54.
29. Paul, P.,Munz, C., 2016. Autophagy and mammalian viruses: roles in immune response, viral replication, and beyond. Adv. Virus Res., Vol 95, 95, 149-195.
30. Sakai, T., Toyoda, A., Hashimoto, K.,Maeda, H., 2000. Isolation and characterization of a novel zinc finger gene, ZNF219, and mapping to the human chromosome 14q11 region. DNA Res., 7, 137-141.
31. Sauter, D., 2014. Counteraction of the multifunctional restriction factor tetherin. Front. Microbiol., 5.
32. Shaid, S., Brandts, C.H., Serve, H.,Dikic, I., 2013. Ubiquitination and selective autophagy. Cell Death Differ., 20, 21-30.
33. Stevenson, G.W., Hoang, H., Schwartz, K.J., Burrough, E.R., Sun, D., Madson, D., Cooper, V.L., Pillatzki, A., Gauger, P., Schmitt, B.J., Koster, L.G., Killian, M.L.,Yoon, K.J., 2013. Emergence of in the United States: clinical signs, lesions, and viral genomic sequences. J. Vet. Diagn. Invest., 25, 649-654.
34. Stubbs, L., Sun, Y.,Caetano-Anolles, D., 2011. Function and evolution of C2H2 zinc finger arrays. Subcell. Biochem., 52, 75-94.
35. Sun, D.B., Feng, L., Shi, H.Y., Chen, J.F., Cui, X.C., Chen, H.Y., Liu, S.W., Tong, Y.E., Wang, Y.F.,Tong, G.Z., 2008. Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. Vet. Microbiol., 131, 73-81.
36. Suzuki, T., Terada, Y., Enjuanes, L., Ohashi, S.,Kamitani, W., 2018. S1 subunit of spike protein from a current highly virulent porcine epidemic diarrhea virus is an important determinant of virulence in piglets. Viruses-Basel, 10.
37. Takigawa, Y., Hata, K., Muramatsu, S., Amano, K., Ono, K., Wakabayashi, M., Matsuda, A., Takada, K., Nishimura, R.,Yoneda, T., 2010a. The transcription factor Znf219 regulates chondrocyte differentiation by assembling a transcription factory with Sox9. J. Cell Sci., 123, 3780-3788.
38. Thavorasak, T., Chulanetra, M., Glab-Ampai, K., Teeranitayatarn, K., Songserm, T., Yodsheewan, R., Sae-Lim, N., Lekcharoensuk, P., Sookrung, N.,Chaicumpa, W., 2022. Novel Neutralizing Epitope of PEDV S1 Protein Identified by Igm Monoclonal Antibody. Viruses-Basel, vol. 14.
39. Wicht, O., Li, W.T., Willems, L., Meuleman, T.J., Wubbolts, R.W., Van Kuppeveld, F.J.M., Rottier, P.J.M.,Bosch, B.J., 2014. Proteolytic activation of the porcine epidemic diarrhea coronavirus spike fusion protein by trypsin in cell culture. J. Virol., 88, 7952-7961.
40. Wood, E.N., 1977. An apparently new syndrome of porcine epidemic diarrhoea. Vet. Rec., 100, 243-244.
41. Yuan, P., Wang, S., Du, T., Liu, L., Chen, X., Yan, J., Han, S., Peng, B., He, X.,Liu, W., 2023. ZNF219, a novel transcriptional repressor, inhibits transcription of the prototype foamy virus by interacting with the viral LTR promoter. Virus Res., 334, 199161.
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10.1016j.virs.2026.03.007-ESM.docx