AIMP2 restricts EV71 replication by recruiting SMURF2 to promote the degradation of 3D polymerase

Junrui Ren; Lei Yu; Qiuhan Zhang; Pengyu Ren; Yumeng Cai; Xueyun Wang; Ke Lan; Shuwen Wu

doi:10.1016/j.virs.2024.06.009

August 2024

Junrui Ren, Lei Yu, Qiuhan Zhang, Pengyu Ren, Yumeng Cai, Xueyun Wang, Ke Lan and Shuwen Wu. AIMP2 restricts EV71 replication by recruiting SMURF2 to promote the degradation of 3D polymerase[J]. Virologica Sinica, 2024, 39(4): 632-644. doi: 10.1016/j.virs.2024.06.009

Citation: Junrui Ren, Lei Yu, Qiuhan Zhang, Pengyu Ren, Yumeng Cai, Xueyun Wang, Ke Lan, Shuwen Wu. AIMP2 restricts EV71 replication by recruiting SMURF2 to promote the degradation of 3D polymerase .VIROLOGICA SINICA, 2024, 39(4) : 632-644. http://dx.doi.org/10.1016/j.virs.2024.06.009

AIMP2招募SMURF2促进3D聚合酶降解抑制EV71复制的功能机制

任军瑞 ^a ,
余雷 ^a,b ,
张秋寒 ^a,b ,
任鹏羽 ^a ,
蔡雨萌 ^a ,
王雪云 ^a ,
蓝柯 ^a,b,c,, ,
吴叔文 ^a,,

cstr: 32224.14.j.virs.2024.06.009

a. 武汉大学生命科学学院、病毒学国家重点实验室;
b. 武汉大学医学研究院;
c. 武汉大学泰康生命医学中心

通讯作者： 蓝柯, klan@whu.edu.cn; 吴叔文, shuwenwu@whu.edu.cn
收稿日期： 2024-03-19
录用日期： 2024-06-25

摘要

肠道病毒71型（EV71）为主的手足口病疫情在亚太地区多次暴发，对婴幼儿健康构成严重威胁。因此，对肠道病毒的感染机制和致病性的研究变得越来越重要。3D聚合酶是EV71复制过程中最关键的RNA依赖RNA聚合酶（RdRp），被广泛用作抑制EV71感染的靶点。在这项研究中，我们报道了一种新的宿主蛋白AIMP2，它可以结合3D聚合酶并抑制EV71复制。进一步研究表明，AIMP2募集E3连接酶SMURF2，介导3D聚合酶的多泛素化和降解。此外，AIMP2的抗病毒作用也适用于CVA16和CVB1血清型。在这项工作中，我们旨在研究AIMP2在EV71感染过程中的动态调控功能，揭示一种新的抗病毒机制，并为开发抗肠道病毒治疗策略提供新的见解。
- AIMP2
- , 肠道病毒71型
- , 3D聚合酶
- , E3连接酶SMURF2
- , 泛素化

AIMP2 restricts EV71 replication by recruiting SMURF2 to promote the degradation of 3D polymerase

Junrui Ren ^a ,
Lei Yu ^a,b ,
Qiuhan Zhang ^a,b ,
Pengyu Ren ^a ,
Yumeng Cai ^a ,
Xueyun Wang ^a ,
Ke Lan ^a,b,c,, ,
Shuwen Wu ^a,,

a. State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China;
b. Medical Research Institute, Wuhan University, Wuhan, 430072, China;
c. Taikang Center for Life and Medical Sciences, Wuhan University, Wuhan, 430072, China

Corresponding author: Ke Lan, klan@whu.edu.cn Shuwen Wu, shuwenwu@whu.edu.cn
Received Date: 19 March 2024
Accepted Date: 25 June 2024

Abstract

Hand, foot and mouth disease (HFMD), mainly caused by enterovirus 71 (EV71), has frequently occurred in the Asia-Pacific region, posing a significant threat to the health of infants and young children. Therefore, research on the infection mechanism and pathogenicity of enteroviruses is increasingly becoming important. The 3D polymerase, as the most critical RNA-dependent RNA polymerase (RdRp) for EV71 replication, is widely targeted to inhibit EV71 infection. In this study, we identified a novel host protein, AIMP2, capable of binding to 3D polymerase and inhibiting EV71 infection. Subsequent investigations revealed that AIMP2 recruits the E3 ligase SMURF2, which mediates the polyubiquitination and degradation of 3D polymerase. Furthermore, the antiviral effect of AIMP2 extended to the CVA16 and CVB1 serotypes. Our research has uncovered the dynamic regulatory function of AIMP2 during EV71 infection, revealing a novel antiviral mechanism and providing new insights for the development of antienteroviral therapeutic strategies.
- AIMP2
- , Enterovirus 71 (EV71)
- , 3D polymerase
- , E3 ligase SMURF2
- , Ubiquitination

References
1. Ang, P.Y., Chong, C.W.H., Alonso, S., 2021. Viral determinants that drive Enterovirus-A71 fitness and virulence. Emerg. Microbes Infect. 10, 713-724.
2. Baer, A., Kehn-Hall, K., 2014. Viral concentration determination through plaque assays: using traditional and novel overlay systems. J. Vis. Exp., e52065.
3. Baggen, J., Thibaut, H.J., Strating, J., van Kuppeveld, F.J.M., 2018. The life cycle of non-polio enteroviruses and how to target it. Nat. Rev. Microbiol. 16, 368-381.
4. Chen, S.C., Chang, L.Y., Wang, Y.W., Chen, Y.C., Weng, K.F., Shih, S.R., Shih, H.M., 2011. Sumoylation-promoted enterovirus 71 3C degradation correlates with a reduction in viral replication and cell apoptosis. J. Biol. Chem. 286, 31373-31384.
5. Choi, J.W., Kim, D.G., Park, M.C., Um, J.Y., Han, J.M., Park, S.G., Choi, E.C., Kim, S., 2009a. AIMP2 promotes TNFalpha-dependent apoptosis via ubiquitin-mediated degradation of TRAF2. J. Cell Sci. 122, 2710-2715.
6. Choi, J.W., Um, J.Y., Kundu, J.K., Surh, Y.J., Kim, S., 2009b. Multidirectional tumor-suppressive activity of AIMP2/p38 and the enhanced susceptibility of AIMP2 heterozygous mice to carcinogenesis. Carcinogenesis 30, 1638-1644.
7. Cox, B., Levent, F., 2018. Hand, foot, and mouth disease. JAMA 320, 2492.
8. Gao, S., Wu, J., Liu, R.Y., Li, J., Song, L., Teng, Y., Sheng, C., Liu, D., Yao, C., Chen, H., Jiang, W., Chen, S., Huang, W., 2015. Interaction of NS2 with AIMP2 facilitates the switch from ubiquitination to SUMOylation of M1 in influenza A virus-infected cells. J. Virol. 89, 300-311.
9. Guo, H., Li, Y., Liu, G., Jiang, Y., Shen, S., Bi, R., Huang, H., Cheng, T., Wang, C., Wei, W., 2019. A second open reading frame in human enterovirus determines viral replication in intestinal epithelial cells. Nat. Commun. 10, 4066.
10. Hagiwara, A., Yoneyama, T., Takami, S., Hashimoto, I., 1984. Genetic and phenotypic characteristics of enterovirus 71 isolates from patients with encephalitis and with hand, foot and mouth disease. Arch. Virol. 79, 273-283.
11. Han, J.M., Park, B.J., Park, S.G., Oh, Y.S., Choi, S.J., Lee, S.W., Hwang, S.K., Chang, S.H., Cho, M.H., Kim, S., 2008. AIMP2/p38, the scaffold for the multi-tRNA synthetase complex, responds to genotoxic stresses via p53. Proc. Natl. Acad. Sci. USA. 105, 11206-11211.
12. Hao, H., Liu, W., Miao, Y., Ma, L., Yu, B., Liu, L., Yang, C., Zhang, K., Chen, Z., Yang, J., Zheng, Z., Zhang, B., Deng, F., Gong, P., Yuan, J., Hu, Z., Guan, W., 2022. N4-acetylcytidine regulates the replication and pathogenicity of enterovirus 71. Nucleic Acids Res. 50, 9339-9354.
13. Hu, Y., Jiang, L., Peng, H.L., 2015. Clinical analysis of 134 children with nervous system damage caused by enterovirus 71 infection. Pediatr. Infect. Dis. J. 34, 718-723.
14. Karim, M., Biquand, E., Declercq, M., Jacob, Y., van der Werf, S., Demeret, C., 2020. Nonproteolytic K29-linked ubiquitination of the PB2 replication protein of influenza A viruses by proviral cullin 4-based E3 ligases. mBio 11 e00305-e00320.
15. Kim, D.G., Lee, J.Y., Lee, J.H., Cho, H.Y., Kang, B.S., Jang, S.Y., Kim, M.H., Guo, M., Han, J.M., Kim, S.J., Kim, S., 2016. Oncogenic mutation of AIMP2/p38 inhibits its tumor-suppressive interaction with Smurf2. Cancer Res. 76, 3422-3436.
16. Kim, S.S., Hur, S.Y., Kim, Y.R., Yoo, N.J., Lee, S.H., 2011. Expression of AIMP1, 2 and 3, the scaffolds for the multi-tRNA synthetase complex, is downregulated in gastric and colorectal cancer. Tumori 97, 380-385.
17. Kumar, R., Mehta, D., Mishra, N., Nayak, D., Sunil, S., 2020. Role of host-mediated post-translational modifications (PTMs) in RNA virus pathogenesis. Int. J. Mol. Sci. 22, 323.
18. Lei, X., Zhang, Z., Xiao, X., Qi, J., He, B., Wang, J., 2017. Enterovirus 71 inhibits pyroptosis through cleavage of gasdermin D. J. Virol. 91, e01069-17.
19. Li, M.L., Brewer, G., 2020. Functional analyses of mammalian virus 5'UTR-derived, small RNAs that regulate virus translation. Methods 183, 13-20.
20. Lin, J.Y., Li, M.L., Shih, S.R., 2009. Far upstream element binding protein 2 interacts with enterovirus 71 internal ribosomal entry site and negatively regulates viral translation. Nucleic Acids Res. 37, 47-59.
21. Liu, X., Xu, Z., Liang, J., Yu, L., Ren, P., Zhou, H.B., Wu, S., Lan, K., 2023. Identification of a novel acylthiourea-based potent broad-spectrum inhibitor for enterovirus 3D polymerase in vitro and in vivo. Antiviral Res. 213, 105583.
22. Liu, Y., Zheng, Z., Shu, B., Meng, J., Zhang, Y., Zheng, C., Ke, X., Gong, P., Hu, Q., Wang, H., 2016. SUMO modification stabilizes enterovirus 71 polymerase 3D to facilitate viral replication. J. Virol. 90, 10472-10485.
23. Lulla, V., Dinan, A.M., Hosmillo, M., Chaudhry, Y., Sherry, L., Irigoyen, N., Nayak, K.M., Stonehouse, N.J., Zilbauer, M., Goodfellow, I., Firth, A.E., 2019. An upstream protein-coding region in enteroviruses modulates virus infection in gut epithelial cells. Nat. Microbiol. 4, 280-292.
24. Nechushtan, H., Kim, S., Kay, G., Razin, E., 2009. Chapter 1: The physiological role of lysyl tRNA synthetase in the immune system. Adv. Immunol. 103, 1-27.
25. Ofir-Birin, Y., Fang, P., Bennett, S.P., Zhang, H.M., Wang, J., Rachmin, I., Shapiro, R., Song, J., Dagan, A., Pozo, J., Kim, S., Marshall, A.G., Schimmel, P., Yang, X.L., Nechushtan, H., Razin, E., Guo, M., 2013. Structural switch of lysyl-tRNA synthetase between translation and transcription. Mol. Cell 49, 30-42.
26. Ooi, M.H., Wong, S.C., Lewthwaite, P., Cardosa, M.J., Solomon, T., 2010. Clinical features, diagnosis, and management of enterovirus 71. Lancet Neurol., 1097-1105.
27. Rivett, A.J., 1990. Eukaryotic protein degradation. Curr. Opin. Cell Biol. 2, 1143-1149.
28. Saguil, A., Kane, S.F., Lauters, R., Mercado, M.G., 2019. Hand-Foot-and-Mouth disease: rapid evidence review. Am. Fam. Physician. 100, 408-414.
29. Schmidt, N.J., Lennette, E.H., Ho, H.H., 1974. An apparently new enterovirus isolated from patients with disease of the central nervous system. J. Infect. Dis. 129, 304-309.
30. Shang, B., Deng, C., Ye, H., Xu, W., Yuan, Z., Shi, P.Y., Zhang, B., 2013. Development and characterization of a stable eGFP enterovirus 71 for antiviral screening. Antiviral Res. 97, 198-205.
31. Shih, S.R., Stollar, V., Li, M.L., 2011. Host factors in enterovirus 71 replication. J. Virol. 85, 9658-9666.
32. Shin, J.Y., Lee, B., Ham, S., Kim, J.H., Kim, H., Kim, H., Jo, M.G., Kim, H.J., Park, S.W., Kweon, H.S., Kim, Y.J., Yun, S.P., Lee, Y., 2022. Pharmacological inhibition of AIMP2 aggregation attenuates α-synuclein aggregation and toxicity in Parkinson's disease. Biomed. Pharmacother. 156, 113908.
33. Solomon, T., Lewthwaite, P., Perera, D., Cardosa, M.J., McMinn, P., Ooi, M.H., 2010. Virology, epidemiology, pathogenesis, and control of enterovirus 71. Lancet Infect. Dis. 10, 778-790.
34. Tang, Q., Li, S., Du, L., Chen, S., Gao, J., Cai, Y., Xu, Z., Zhao, Z., Lan, K., Wu, S., 2020. Emetine protects mice from enterovirus infection by inhibiting viral translation. Antiviral Res. 173, 104650.
35. Wang, H., Li, Y., 2019. Recent progress on functional genomics research of enterovirus 71. Virol. Sin. 34, 9-21.
36. Wang, K., Zou, C., Wang, X., Huang, C., Feng, T., Pan, W., Wu, Q., Wang, P., Dai, J., 2018. Interferon-stimulated TRIM69 interrupts dengue virus replication by ubiquitinating viral nonstructural protein 3. PLoS Pathog. 14, e1007287.
37. Wang, X., Hu, Z., Zhang, W., Wu, S., Hao, Y., Xiao, X., Li, J., Yu, X., Yang, C., Wang, J., Zhang, H., Ma, F., Shi, W., Wang, J., Lei, X., Zhang, X., He, S., 2023. Inhibition of lysosome-tethered Ragulator-Rag-3D complex restricts the replication of Enterovirus 71 and Coxsackie A16. J. Cell Biol. 222, e202303108.
38. Xia, H., Wang, P., Wang, G.C., Yang, J., Sun, X., Wu, W., Qiu, Y., Shu, T., Zhao, X., Yin, L., Qin, C.F., Hu, Y., Zhou, X., 2015. Human enterovirus nonstructural protein 2CATPase functions as both an RNA helicase and ATP-independent RNA chaperone. PLoS Pathog. 11, e1005067.
39. Xiao, H., Li, J., Yang, X., Li, Z., Wang, Y., Rui, Y., Liu, B., Zhang, W., 2021. Ectopic expression of TRIM25 restores RIG-I expression and IFN production reduced by multiple enteroviruses 3C(pro). Virol. Sin. 36, 1363-1374.
40. You, Q., Wu, J., Liu, Y., Zhang, F., Jiang, N., Tian, X., Cai, Y., Yang, E., Lyu, R., Zheng, N., Chen, D., Wu, Z., 2023. HMGB1 release induced by EV71 infection exacerbates blood-brain barrier disruption via VE-cadherin phosphorylation. Virus Res. 338, 199240.
41. Yuan, J., Shen, L., Wu, J., Zou, X., Gu, J., Chen, J., Mao, L., 2018. Enterovirus A71 proteins: structure and function. Front. Microbiol. 9, 286.
42. Yum, M.K., Kang, J.S., Lee, A.E., Jo, Y.W., Seo, J.Y., Kim, H.A., Kim, Y.Y., Seong, J., Lee, E.B., Kim, J.H., Han, J.M., Kim, S., Kong, Y.Y., 2016. AIMP2 controls intestinal stem cell compartments and tumorigenesis by modulating Wnt/β-catenin signaling. Cancer Res. 76, 4559-4568.
43. Zhang, H., Song, L., Cong, H., Tien, P., 2015. Nuclear protein Sam68 interacts with the enterovirus 71 internal ribosome entry site and positively regulates viral protein translation. J. Virol. 89, 10031-10043.
44. Zhang, Q., Li, S., Lei, P., Li, Z., Chen, F., Chen, Q., Wang, Y., Gong, J., Tang, Q., Liu, X., Lan, K., Wu, S., 2021. ANXA2 facilitates enterovirus 71 infection by interacting with 3D polymerase and PI4KB to assist the assembly of replication organelles. Virol. Sin. 36, 1387-1399.
45. Zhou, X., Tian, L., Wang, J., Zheng, B., Zhang, W., 2022. EV71 3C protease cleaves host anti-viral factor OAS3 and enhances virus replication. Virol. Sin. 37, 418-426.
46. Zhou, Z., Sun, B., Huang, S., Yu, D., Zhang, X., 2020a. Roles of aminoacyl-tRNA synthetase-interacting multi-functional proteins in physiology and cancer. Cell Death Dis. 11, 579.
47. Zhou, Z., Sun, B., Nie, A., Yu, D., Bian, M., 2020b. Roles of aminoacyl-tRNA synthetases in cancer. Front. Cell Dev. Biol. 8, 599765.
48. Zhu, Q., Yu, T., Gan, S., Wang, Y., Pei, Y., Zhao, Q., Pei, S., Hao, S., Yuan, J., Xu, J., Hou, F., Wu, X., Peng, C., Wu, P., Qin, J., Xiao, Y., 2020. TRIM24 facilitates antiviral immunity through mediating K63-linked TRAF3 ubiquitination. J. Exp. Med. 217, e20192083.
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