Citation: Huimin Xiao, Jingliang Li, Xu Yang, Zhaolong Li, Ying Wang, Yajuan Rui, Bin Liu, Wenyan Zhang. Ectopic Expression of TRIM25 Restores RIG-I Expression and IFN Production Reduced by Multiple Enteroviruses 3Cpro .VIROLOGICA SINICA, 2021, 36(6) : 1363-1374.  http://dx.doi.org/10.1007/s12250-021-00410-x

Ectopic Expression of TRIM25 Restores RIG-I Expression and IFN Production Reduced by Multiple Enteroviruses 3Cpro

  • Corresponding author: Bin Liu, kjkliubin@163.com, ORCID: http://orcid.org/0000-0001-8658-9888
    Wenyan Zhang, zhangwenyan@jlu.edu.cn, ORCID: http://orcid.org/0000-0003-4507-521X
  • Received Date: 31 January 2021
    Accepted Date: 12 April 2021
    Published Date: 25 June 2021
    Available online: 01 December 2021
  • Enteroviruses (EVs) 3C proteins suppress type Ⅰ interferon (IFN) responses mediated by retinoid acid-inducible gene I (RIG-I), while an E3 ubiquitin ligase, tripartite motif protein 25 (TRIM25)-mediated RIG-I ubiquitination is essential for RIG-I antiviral activity. Therefore, whether the effect of EVs 3C on RIG-I is associated with TRIM25 expression is worth to be further investigated. Here, we demonstrate that 3C proteins of EV71 and coxsackievirus B3 (CVB3) reduced not only RIG-I expression but also TRIM25 expression through protease cleavage activity, while overexpression of TRIM25 restored RIG-I expression and IFN-β production reduced by 3C proteins. Further investigation confirmed that the two amino acids and functional domains in TRIM25 required for RIG-I ubiquitination and TRIM25 structural conformation were essential for the recovery of RIG-I expression. Moreover, we also observed that TRIM25 could rescue RIG-I expression reduced by 3C proteins of CVA6 and EV-D68 but not CVA16. Our findings provide an insightful interpretation of 3C-mediated host innate immune suppression and support TRIM25 as an attractive target against multiple EVs infection.


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    1. Barral PM, Morrison JM, Drahos J, Gupta P, Sarkar D, Fisher PB, Racaniello VR (2007) MDA-5 is cleaved in poliovirus-infected cells. J Virol 81: 3677-3684
        doi: 10.1128/JVI.01360-06

    2. Barral PM, Sarkar D, Fisher PB, Racaniello VR (2009) RIG-I is cleaved during picornavirus infection. Virology 391: 171-176
        doi: 10.1016/j.virol.2009.06.045

    3. Castanier C, Zemirli N, Portier A, Garcin D, Bidere N, Vazquez A, Arnoult D (2012) MAVS ubiquitination by the E3 ligase TRIM25 and degradation by the proteasome is involved in type Ⅰ interferon production after activation of the antiviral RIG-I-like receptors. BMC Biol 10: 44
        doi: 10.1186/1741-7007-10-44

    4. Croft SN, Walker EJ, Ghildyal R (2017) Picornaviruses and apoptosis: subversion of cell death. MBio 8: e01009-17
        doi: 10.1128/mBio.01009-17

    5. de Breyne S, Bonderoff JM, Chumakov KM, Lloyd RE, Hellen CU (2008) Cleavage of eukaryotic initiation factor eIF5B by enterovirus 3C proteases. Virology 378: 118-122
        doi: 10.1016/j.virol.2008.05.019

    6. Etchison D, Milburn SC, Edery I, Sonenberg N, Hershey JW (1982) Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220, 000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. J Biol Chem 257: 14806-14810
        doi: 10.1016/S0021-9258(18)33352-0

    7. Feng Q, Langereis MA, Lork M, Nguyen M, Hato SV, Lanke K, Emdad L, Bhoopathi P, Fisher PB, Lloyd RE, van Kuppeveld FJ (2014) Enterovirus 2Apro targets MDA5 and MAVS in infected cells. J Virol 88: 3369-3378
        doi: 10.1128/JVI.02712-13

    8. Gack MU, Shin YC, Joo CH, Urano T, Liang C, Sun L, Takeuchi O, Akira S, Chen Z, Inoue S, Jung JU (2007) TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature 446: 916-920
        doi: 10.1038/nature05732

    9. Gack MU, Kirchhofer A, Shin YC, Inn KS, Liang C, Cui S, Myong S, Ha T, Hopfner KP, Jung JU (2008) Roles of RIG-I N-terminal tandem CARD and splice variant in TRIM25-mediated antiviral signal transduction. Proc Natl Acad Sci U S A 105: 16743-16748
        doi: 10.1073/pnas.0804947105

    10. Gack MU, Albrecht RA, Urano T, Inn KS, Huang IC, Carnero E, Farzan M, Inoue S, Jung JU, Garcia-Sastre A (2009) Influenza A virus NS1 targets the ubiquitin ligase TRIM25 to evade recognition by the host viral RNA sensor RIG-I. Cell Host Microbe 5: 439-449
        doi: 10.1016/j.chom.2009.04.006

    11. Hu Y, Li W, Gao T, Cui Y, Jin Y, Li P, Ma Q, Liu X, Cao C (2017) The severe acute respiratory syndrome coronavirus nucleocapsid inhibits type Ⅰ interferon production by interfering with TRIM25-mediated RIG-I ubiquitination. J Virol 91: e02143-16

    12. Hung HC, Wang HC, Shih SR, Teng IF, Tseng CP, Hsu JT (2011) Synergistic inhibition of enterovirus 71 replication by interferon and rupintrivir. J Infect Dis 203: 1784-1790
        doi: 10.1093/infdis/jir174

    13. Kuyumcu-Martinez NM, Van Eden ME, Younan P, Lloyd RE (2004) Cleavage of poly(A)-binding protein by poliovirus 3C protease inhibits host cell translation: a novel mechanism for host translation shutoff. Mol Cell Biol 24: 1779-1790
        doi: 10.1128/MCB.24.4.1779-1790.2004

    14. Lee YP, Wang YF, Wang JR, Huang SW, Yu CK (2012) Enterovirus 71 blocks selectively type Ⅰ interferon production through the 3C viral protein in mice. J Med Virol 84: 1779-1789
        doi: 10.1002/jmv.23377

    15. Lee NR, Kim HI, Choi MS, Yi CM, Inn KS (2015) Regulation of MDA5-MAVS antiviral signaling axis by TRIM25 through TRAF6-mediated NF-kappaB activation. Mol Cells 38: 759-764
        doi: 10.14348/molcells.2015.0047

    16. Lei X, Liu X, Ma Y, Sun Z, Yang Y, Jin Q, He B, Wang J (2010) The 3C protein of enterovirus 71 inhibits retinoid acid-inducible gene I-mediated interferon regulatory factor 3 activation and type Ⅰ interferon responses. J Virol 84: 8051-8061
        doi: 10.1128/JVI.02491-09

    17. Lei X, Sun Z, Liu X, Jin Q, He B, Wang J (2011) Cleavage of the adaptor protein TRIF by enterovirus 71 3C inhibits antiviral responses mediated by Toll-like receptor 3. J Virol 85: 8811-8818
        doi: 10.1128/JVI.00447-11

    18. 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
        doi: 10.1128/JVI.01855-12

    19. Lei X, Han N, Xiao X, Jin Q, He B, Wang J (2014) Enterovirus 71 3C inhibits cytokine expression through cleavage of the TAK1/TAB1/TAB2/TAB3 complex. J Virol 88: 9830-9841
        doi: 10.1128/JVI.01425-14

    20. Li ML, Hsu TA, Chen TC, Chang SC, Lee JC, Chen CC, Stollar V, Shih SR (2002) The 3C protease activity of enterovirus 71 induces human neural cell apoptosis. Virology 293: 386-395
        doi: 10.1006/viro.2001.1310

    21. Li J, Yao Y, Chen Y, Xu X, Lin Y, Yang Z, Qiao W, Tan J (2017) Enterovirus 71 3C promotes apoptosis through cleavage of PinX1, a telomere binding protein. J Virol 91: e02016-16
        doi: 10.1128/JVI.02016-16

    22. Li ML, Lin JY, Chen BS, Weng KF, Shih SR, Calderon JD, Tolbert BS, Brewer G (2019) EV71 3C protease induces apoptosis by cleavage of hnRNP A1 to promote apaf-1 translation. PLoS ONE 14: e0221048
        doi: 10.1371/journal.pone.0221048

    23. McMinn PC (2002) An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS Microbiol Rev 26: 91-107
        doi: 10.1111/j.1574-6976.2002.tb00601.x

    24. Mukherjee A, Morosky SA, Delorme-Axford E, Dybdahl-Sissoko N, Oberste MS, Wang T, Coyne CB (2011) The coxsackievirus B 3C protease cleaves MAVS and TRIF to attenuate host type Ⅰ interferon and apoptotic signaling. PLoS Pathog 7: e1001311
        doi: 10.1371/journal.ppat.1001311

    25. Munir M (2010) TRIM proteins: another class of viral victims. Sci Signal 3: jc2

    26. Nisole S, Stoye JP, Saib A (2005) TRIM family proteins: retroviral restriction and antiviral defence. Nat Rev Microbiol 3: 799-808
        doi: 10.1038/nrmicro1248

    27. Ozato K, Shin DM, Chang TH, Morse HC 3rd (2008) TRIM family proteins and their emerging roles in innate immunity. Nat Rev Immunol 8: 849-860
        doi: 10.1038/nri2413

    28. Rajsbaum R, Albrecht RA, Wang MK, Maharaj NP, Versteeg GA, Nistal-Villan E, Garcia-Sastre A, Gack MU (2012) Species-specific inhibition of RIG-I ubiquitination and IFN induction by the influenza A virus NS1 protein. PLoS Pathog 8: e1003059
        doi: 10.1371/journal.ppat.1003059

    29. Rui Y, Su J, Wang H, Chang J, Wang S, Zheng W, Cai Y, Wei W, Gordy JT, Markham R, Kong W, Zhang W, Yu XF (2017) Disruption of MDA5-mediated innate immune responses by the 3C proteins of coxsackievirus A16, coxsackievirus A6, and enterovirus D68. J Virol 91: e00546-17
        doi: 10.1128/JVI.00546-17

    30. Sanchez JG, Chiang JJ, Sparrer KMJ, Alam SL, Chi M, Roganowicz MD, Sankaran B, Gack MU, Pornillos O (2016) Mechanism of TRIM25 catalytic activation in the antiviral RIG-I pathway. Cell Rep 16: 1315-1325
        doi: 10.1016/j.celrep.2016.06.070

    31. Sumpter R Jr, Loo YM, Foy E, Li K, Yoneyama M, Fujita T, Lemon SM, Gale M Jr (2005) Regulating intracellular antiviral defense and permissiveness to hepatitis C virus RNA replication through a cellular RNA helicase, RIG-I. J Virol 79: 2689-2699
        doi: 10.1128/JVI.79.5.2689-2699.2005

    32. Wang B, Xi X, Lei X, Zhang X, Cui S, Wang J, Jin Q, Zhao Z (2013) Enterovirus 71 protease 2Apro targets MAVS to inhibit anti-viral type Ⅰ interferon responses. PLoS Pathog 9: e1003231
        doi: 10.1371/journal.ppat.1003231

    33. Wang H, Lei X, Xiao X, Yang C, Lu W, Huang Z, Leng Q, Jin Q, He B, Meng G, Wang J (2015) Reciprocal regulation between enterovirus 71 and the NLRP3 inflammasome. Cell Rep 12: 42-48
        doi: 10.1016/j.celrep.2015.05.047

    34. Xiang Z, Liu L, Lei X, Zhou Z, He B, Wang J (2015) 3C Protease of enterovirus D68 inhibits cellular defense mediated by interferon regulatory factor 7. J Virol 90: 1613-1621
        doi: 10.1128/JVI.02395-15

    35. Xiao J, Zhao Y, Wang H, Yuan Y, Yang F, Zhang C, Kai G (2011) Non-covalent interaction of dietary polyphenols with total plasma proteins of type Ⅱ diabetes: molecular structure/property-affinity relationships. Integr Biol 3: 1087-1094
        doi: 10.1039/c1ib00076d

    36. Zhu Q, Hao Y, Ma J, Yu S, Wang Y (2011) Surveillance of hand, foot, and mouth disease in mainland China (2008-2009). Biomed Environ Sci 24: 349-356

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    Ectopic Expression of TRIM25 Restores RIG-I Expression and IFN Production Reduced by Multiple Enteroviruses 3Cpro

      Corresponding author: Bin Liu, kjkliubin@163.com
      Corresponding author: Wenyan Zhang, zhangwenyan@jlu.edu.cn
    • 1. Institute of Virology and AIDS Research, Key Laboratory of Organ Regeneration and Transplantation of The Ministry of Education, The First Hospital of Jilin University, Changchun, 130021, China
    • 2. Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, Ministry of Education), Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310058, China
    • 3. Department of Hand Surgery, First Hospital of Jilin University, Changchun, 130021, China
    • 4. Changchun Institute of Biological Products Co., Ltd, Changchun, 130012, China

    Abstract: 

    Enteroviruses (EVs) 3C proteins suppress type Ⅰ interferon (IFN) responses mediated by retinoid acid-inducible gene I (RIG-I), while an E3 ubiquitin ligase, tripartite motif protein 25 (TRIM25)-mediated RIG-I ubiquitination is essential for RIG-I antiviral activity. Therefore, whether the effect of EVs 3C on RIG-I is associated with TRIM25 expression is worth to be further investigated. Here, we demonstrate that 3C proteins of EV71 and coxsackievirus B3 (CVB3) reduced not only RIG-I expression but also TRIM25 expression through protease cleavage activity, while overexpression of TRIM25 restored RIG-I expression and IFN-β production reduced by 3C proteins. Further investigation confirmed that the two amino acids and functional domains in TRIM25 required for RIG-I ubiquitination and TRIM25 structural conformation were essential for the recovery of RIG-I expression. Moreover, we also observed that TRIM25 could rescue RIG-I expression reduced by 3C proteins of CVA6 and EV-D68 but not CVA16. Our findings provide an insightful interpretation of 3C-mediated host innate immune suppression and support TRIM25 as an attractive target against multiple EVs infection.