For best viewing of the website please use Mozilla Firefox or Google Chrome.

. doi: 10.1016/j.virs.2022.04.013
Citation: Xiaolei Zhou, Li Tian, Jian Wang, Baisong Zheng, Wenyan Zhang. EV71 3C protease cleaves host anti-viral factor OAS3 and enhances virus replication [J].VIROLOGICA SINICA, 2022, 37(3) : 418-426.  http://dx.doi.org/10.1016/j.virs.2022.04.013

EV71通过3C蛋白酶切割宿主抗病毒因子OAS3,促进病毒复制

  • 肠道病毒的全球传播变得更加频繁,严重威胁生命。I型干扰素已被证明可通过调节干扰素刺激基因的表达来控制肠道病毒。2'-5'-寡腺苷酸合成酶3(OAS3)是OAS/RNase L抗病毒系统中的一个重要的干扰素刺激基因。然而,OAS3和肠道病毒之间的关系尚不清楚。在这里,我们揭示了OAS3相对于OAS1和OAS2,在体外具有显著抑制肠道病毒EV71复制的能力。然而,EV71可以利用自体的3C蛋白酶(3Cpro)切割细胞内的OAS3,从而增强病毒的复制。研究发现人类鼻病毒3C蛋白酶的抑制剂Rupintrivir可以完全消除了EV71 3Cpro对OAS3的切割。EV71 3Cpro的蛋白水解缺陷突变体H40G、E71A和C147G也丧失了切割OAS3的能力。机制研究表明,OAS3蛋白C末端的Q982-G983基序被确定为EV71 3Cpro切割的一个关键位点。进一步的研究表明,OAS3不仅能抑制EV71,还能抑制柯萨奇病毒B3(CVB3)、柯萨奇病毒A16(CA16)、肠病毒D68(EVD68)和柯萨奇病毒A16(CA6)亚型。值得注意的是,与其他四种亚型病毒不同,CA16 3Cpro不能切割OAS3。CA16 3Cpro中的两个关键氨基酸Ile36和Val86的变异,可能导致其切割OAS和3AB蛋白能力减弱,而使CA16病毒复制减弱和延迟。我们的工作阐明了OAS3广泛的抗肠道病毒功能,并阐明了EV71使用3Cpro逃避OAS3抗病毒作用的新机制。这些发现可以作为开发抗肠道病毒新疗法的重要切入点。

EV71 3C protease cleaves host anti-viral factor OAS3 and enhances virus replication

  • The global spread of enteroviruses (EVs) has become more frequent, severe and life-threatening. Intereron (IFN) I has been proved to control EVs by regulating IFN-stimulated genes (ISG) expression. 20-50-oligoadenylate synthetases 3 (OAS3) is an important ISG in the OAS/RNase L antiviral system. The relationship between OAS3 and EVs is still unclear. Here, we reveal that OAS3, superior to OAS1 and OAS2, significantly inhibited EV71 replication in vitro. However, EV71 utilized autologous 3C protease (3Cpro) to cleave intracellular OAS3 and enhance viral replication. Rupintrivir, a human rhinovirus 3C protease inhibitor, completely abolished the cleavage of EV71 3Cpro on OAS3. And the proteolytically deficient mutants H40G, E71A, and C147G of EV71 3Cpro also lost the ability of OAS3 cleavage. Mechanistically, the Q982-G983 motif in C-terminal of OAS3 was identified as a crucial 3Cpro cutting site. Further investigation indicated that OAS3 inhibited not only EV71 but also Coxsackievirus B3 (CVB3), Coxsackievirus A16 (CA16), Enterovirus D68 (EVD68), and Coxsackievirus A6 (CA6) subtypes. Notably, unlike other four subtypes, CA16 3Cpro could not cleave OAS3. Two key amino acids variation Ile36 and Val86 in CA16 3Cpro might result in weak and delayed virus replication of CA16 because of failure of OAS and 3AB cleavage. Our works elucidate the broad anti-EVs function of OAS3, and illuminate a novel mechanism by which EV71 use 3Cpro to escape the antiviral effect of OAS3. These findings can be an important entry point for developing novel therapeutic strategies for multiple EVs infection.

  • 加载中
    1. Baggen, J., Thibaut, H.J., Strating, J., van Kuppeveld, F.J.M., 2018. The life cycle of nonpolio enteroviruses and how to target it. Nat. Rev. Microbiol. 16, 368-381.

    2. Chang, H.W., Watson, J.C., Jacobs, B.L., 1992. The e3l gene of vaccinia virus encodes an inhibitor of the interferon-induced, double-stranded RNA-dependent protein kinase.Proc. Natl. Acad. Sci. Unit. States Am. 89, 4825-4829.

    3. Coccia, E.M., Romeo, G., Nissim, A., Marziali, G., Albertini, R., Affabris, E., Battistini, A., Fiorucci, G., Orsatti, R., Rossi, G.B., et al., 1990. A full-length murine 2-5a synthetase cdna transfected in nih-3t3 cells impairs emcv but not vsv replication. Virology 179, 228-233.

    4. Ghosh, A., Sarkar, S.N., Sen, G.C., 2000. Cell growth regulatory and antiviral effects of the p69 isozyme of 2-5 (a) synthetase. Virology 266, 319-328.

    5. Han, J.Q., Townsend, H.L., Jha, B.K., Paranjape, J.M., Silverman, R.H., Barton, D.J., 2007. A phylogenetically conserved RNA structure in the poliovirus open reading frame inhibits the antiviral endoribonuclease Rnase l. J. Virol. 81, 5561-5572.

    6. Hovanessian, A.G., Justesen, J., 2007. The human 2'-5'oligoadenylate synthetase family:unique interferon-inducible enzymes catalyzing 2'-5' instead of 3'-5' phosphodiester bond formation. Biochimie 89, 779-788.

    7. Huismans, H., Joklik, W.K., 1976. Reovirus-coded polypeptides in infected cells:isolation of two native monomeric polypeptides with affinity for single-stranded and doublestranded RNA, respectively. Virology 70, 411-424.

    8. Ibsen, M.S., Gad, H.H., Andersen, L.L., Hornung, V., Julkunen, I., Sarkar, S.N., Hartmann, R., 2015. Structural and functional analysis reveals that human OASL binds dsRNA to enhance RIG-I signaling. Nucleic Acids Res. 43, 5236-5248.

    9. Justesen, J., Hartmann, R., Kjeldgaard, N.O., 2000. Gene structure and function of the 2'-5'-oligoadenylate synthetase family. Cell. Mol. Life Sci. 57, 1593-1612.

    10. Kitamura, N., Semler, B.L., Rothberg, P.G., Larsen, G.R., Adler, C.J., Dorner, A.J., Emini, E.A., Hanecak, R., Lee, J.J., van der Werf, S., Anderson, C.W., Wimmer, E., 1981. Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature 291, 547-553.

    11. Kristiansen, H., Gad, H.H., Eskildsen-Larsen, S., Despres, P., Hartmann, R., 2011. The oligoadenylate synthetase family:an ancient protein family with multiple antiviral activities. J. Interferon Cytokine Res. 31, 41-47.

    12. Laitinen, O.H., Svedin, E., Kapell, S., Nurminen, A., Hytönen, V.P., Flodström-Tullberg, M., 2016. Enteroviral proteases:structure, host interactions and pathogenicity. Rev. Med. Virol. 26, 251-267.

    13. Lei, X., Xiao, X., Xue, Q., Jin, Q., He, B., Wang, J., 2013. Cleavage of interferon regulatory factor 7 by enterovirus 71 3c suppresses cellular responses. J. Virol. 87, 1690-1698.

    14. 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.

    15. Lin, J.Y., Chen, T.C., Weng, K.F., Chang, S.C., Chen, L.L., Shih, S.R., 2009. Viral and host proteins involved in picornavirus life cycle. J. Biomed. Sci. 16, 103.

    16. Liu, Y., Liu, P., Liu, S., Guo, Y., He, H., Yang, C., Song, J., Zhang, N., Cheng, J., Chen, Z., 2018. Oligoadenylate synthetase 3 s381r gene polymorphism is associated with severity of ev71 infection in Chinese children. J. Clin. Virol. 101, 29-33.

    17. Lu, J., Yi, L., Zhao, J., Yu, J., Chen, Y., Lin, M.C., Kung, H.F., He, M.L., 2012. Enterovirus 71 disrupts interferon signaling by reducing the level of interferon receptor 1.J. Virol. 86, 3767-3776.

    18. Lu, J., Yi, L., Ke, C., Zhang, Y., Liu, R., Chen, J., Kung, H.F., He, M.L., 2015. The interaction between human enteroviruses and type I IFN signaling pathway. Crit.Rev. Microbiol. 41, 201-207.

    19. Min, J.-Y., Krug, R.M., 2006. The primary function of RNA binding by the influenza A virus ns1 protein in infected cells:inhibiting the 20-50 oligo (a) synthetase/RNAse l pathway. Proc. Natl. Acad. Sci. Unit. States Am. 103, 7100-7105.

    20. Muehlenbachs, A., Bhatnagar, J., Zaki, S.R., 2015. Tissue tropism, pathology and pathogenesis of enterovirus infection. J. Pathol. 235, 217-228.

    21. Pollack, A., Kontorovich, A.R., Fuster, V., Dec, G.W., 2015. Viral myocarditis-diagnosis, treatment options, and current controversies. Nat. Rev. Cardiol. 12, 670-680.

    22. Pons-Salort, M., Parker, E.P., Grassly, N.C., 2015. The epidemiology of non-polio enteroviruses:recent advances and outstanding questions. Curr. Opin. Infect. Dis. 28, 479-487.

    23. Rysiecki, G., Gewert, D.R., Williams, B.R., 1989. Constitutive expression of a 2',5'-oligoadenylate synthetase cdna results in increased antiviral activity and growth suppression. J. Interferon Res. 9, 649-657.

    24. Schroder, H.C., Ugarković, D., Wenger, R., Reuter, P., Okamoto, T., Müller, W.E., 1990.Binding of tat protein to tar region of human immunodeficiency virus type 1 blocks tar-mediated activation of (2'-5')oligoadenylate synthetase. AIDS Res. Hum. Retrovir. 6, 659-672.

    25. Silverman, R.H., 2007. Viral encounters with 2',5'-oligoadenylate synthetase and RNase l during the interferon antiviral response. J. Virol. 81, 12720-12729.

    26. 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.

    27. Tan, Y., Yang, T., Liu, P., Chen, L., Tian, Q., Guo, Y., He, H., Liu, Y., Chen, Z., 2017.Association of the oas3 rs1859330 g/a genetic polymorphism with severity of enterovirus-71 infection in Chinese han children. Arch. Virol. 162, 2305-2313.

    28. Wang, J., Fan, T., Yao, X., Wu, Z., Guo, L., Lei, X., Wang, J., Wang, M., Jin, Q., Cui, S., 2011. Crystal structures of enterovirus 71 3c protease complexed with rupintrivir reveal the roles of catalytically important residues. J. Virol. 85, 10021-10030.

    29. Weng, K.F., Li, M.L., Hung, C.T., Shih, S.R., 2009. Enterovirus 71 3c protease cleaves a novel target cstf-64 and inhibits cellular polyadenylation. PLoS Pathog. 5, e1000593.

    30. Xu, N., Yang, J., Zheng, B., Zhang, Y., Cao, Y., Huan, C., Wang, S., Chang, J., Zhang, W., 2020. The pyrimidine analog fnc potently inhibits the replication of multiple enteroviruses. J. Virol. 94.

    31. Zhang, R., Jha, B.K., Ogden, K.M., Dong, B., Zhao, L., Elliott, R., Patton, J.T., Silverman, R.H., Weiss, S.R., 2013. Homologous 20,50-phosphodiesterases from disparate RNA viruses antagonize antiviral innate immunity. Proc. Natl. Acad. Sci.Unit. States Am. 110, 13114-13119.

    32. Zhao, L., Jha Babal, K., Wu, A., Elliott, R., Ziebuhr, J., Gorbalenya Alexander, E., Silverman Robert, H., Weiss Susan, R., 2012. Antagonism of the interferon-induced oas-RNase l pathway by murine coronavirus ns2 protein is required for virus replication and liver pathology. Cell Host Microbe 11, 607-616.

    33. Zhou, A., Hassel, B.A., Silverman, R.H., 1993. Expression cloning of 2-5a-dependent RNAase:a uniquely regulated mediator of interferon action. Cell 72, 753-765.

  • 加载中
  • 10.1016j.virs.2022.04.013-ESM.docx

Article Metrics

Article views(1288) PDF downloads(16) Cited by(0)

Related
Proportional views
    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    EV71 3C protease cleaves host anti-viral factor OAS3 and enhances virus replication

      Corresponding author: Baisong Zheng, zhengbs@jlu.edu.cn
      Corresponding author: Wenyan Zhang, zhangwenyan@jlu.edu.cn
    • Center for Infectious Diseases and Pathogen Biology, Institute of Virology and AIDS Research, Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education, The First Hospital of Jilin University, Jilin, 130021, China

    Abstract: The global spread of enteroviruses (EVs) has become more frequent, severe and life-threatening. Intereron (IFN) I has been proved to control EVs by regulating IFN-stimulated genes (ISG) expression. 20-50-oligoadenylate synthetases 3 (OAS3) is an important ISG in the OAS/RNase L antiviral system. The relationship between OAS3 and EVs is still unclear. Here, we reveal that OAS3, superior to OAS1 and OAS2, significantly inhibited EV71 replication in vitro. However, EV71 utilized autologous 3C protease (3Cpro) to cleave intracellular OAS3 and enhance viral replication. Rupintrivir, a human rhinovirus 3C protease inhibitor, completely abolished the cleavage of EV71 3Cpro on OAS3. And the proteolytically deficient mutants H40G, E71A, and C147G of EV71 3Cpro also lost the ability of OAS3 cleavage. Mechanistically, the Q982-G983 motif in C-terminal of OAS3 was identified as a crucial 3Cpro cutting site. Further investigation indicated that OAS3 inhibited not only EV71 but also Coxsackievirus B3 (CVB3), Coxsackievirus A16 (CA16), Enterovirus D68 (EVD68), and Coxsackievirus A6 (CA6) subtypes. Notably, unlike other four subtypes, CA16 3Cpro could not cleave OAS3. Two key amino acids variation Ile36 and Val86 in CA16 3Cpro might result in weak and delayed virus replication of CA16 because of failure of OAS and 3AB cleavage. Our works elucidate the broad anti-EVs function of OAS3, and illuminate a novel mechanism by which EV71 use 3Cpro to escape the antiviral effect of OAS3. These findings can be an important entry point for developing novel therapeutic strategies for multiple EVs infection.

    Reference (33) Relative (20)

    目录

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return