1887

Abstract

Enterovirus A71 (EV-A71) is a positive-strand RNA virus that causes hand-foot-mouth disease and neurological complications in children and infants. Although the underlying mechanisms remain to be further defined, impaired immunity is thought to play an important role. The host zinc-finger antiviral protein (ZAP), an IFN-stimulated gene product, has been reported to specifically inhibit the replication of certain viruses. However, whether ZAP restricts the infection of enteroviruses remains unknown. Here, we report that EV-A71 infection upregulates ZAP mRNA in RD and HeLa cells. Moreover, ZAP overexpression rendered 293 T cells resistant to EV-A71 infection, whereas siRNA-mediated depletion of endogenous ZAP enhanced EV-A71 infection. The EV-A71 infection stimulated site-specific proteolysis of two ZAP isoforms, leading to the accumulation of a 40 kDa N-terminal ZAP fragment in virus-infected cells. We further revealed that the 3C protease (3Cpro) of EV-A71 mediates ZAP cleavage, which requires protease activity. Furthermore, ZAP variants with single amino acid substitutions at Gln-369 were resistant to 3Cpro cleavage, implying that Gln-369 is the sole cleavage site in ZAP. Moreover, although ZAP overexpression inhibited EV-A71 replication, the cleaved fragments did not show this effect. Our results indicate that an equilibrium between ZAP and enterovirus 3Cpro controls viral infection. The findings in this study suggest that viral 3Cpro mediated ZAP cleavage may represent a mechanism to escape host antiviral responses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000982
2017-11-28
2019-12-05
Loading full text...

Full text loading...

/deliver/fulltext/jgv/99/1/73.html?itemId=/content/journal/jgv/10.1099/jgv.0.000982&mimeType=html&fmt=ahah

References

  1. Solomon T, Lewthwaite P, Perera D, Cardosa MJ, Mcminn P et al. Virology, epidemiology, pathogenesis, and control of enterovirus 71. Lancet Infect Dis 2010;10:778–790 [CrossRef][PubMed]
    [Google Scholar]
  2. Schmidt NJ, Lennette EH, Ho HH. An apparently new enterovirus isolated from patients with disease of the central nervous system. J Infect Dis 1974;129:304–309 [CrossRef][PubMed]
    [Google Scholar]
  3. Mcminn PC. An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS Microbiol Rev 2002;26:91–107 [CrossRef][PubMed]
    [Google Scholar]
  4. Lei X, Han N, Xiao X, Jin Q, He B et al. Enterovirus 71 3C inhibits cytokine expression through cleavage of the TAK1/TAB1/TAB2/TAB3 complex. J Virol 2014;88:9830–9841 [CrossRef][PubMed]
    [Google Scholar]
  5. Lei X, Sun Z, Liu X, Jin Q, He B et al. Cleavage of the adaptor protein TRIF by enterovirus 71 3C inhibits antiviral responses mediated by Toll-like receptor 3. J Virol 2011;85:8811–8818 [CrossRef][PubMed]
    [Google Scholar]
  6. Lei X, Xiao X, Xue Q, Jin Q, He B et al. Cleavage of interferon regulatory factor 7 by enterovirus 71 3C suppresses cellular responses. J Virol 2013;87:1690–1698 [CrossRef][PubMed]
    [Google Scholar]
  7. Li ML, Hsu TA, Chen TC, Chang SC, Lee JC et al. The 3C protease activity of enterovirus 71 induces human neural cell apoptosis. Virology 2002;293:386–395 [CrossRef][PubMed]
    [Google Scholar]
  8. Weng KF, Li ML, Hung CT, Shih SR. Enterovirus 71 3C protease cleaves a novel target CstF-64 and inhibits cellular polyadenylation. PLoS Pathog 2009;5:e1000593 [CrossRef][PubMed]
    [Google Scholar]
  9. Chen LL, Kung YA, Weng KF, Lin JY, Horng JT et al. Enterovirus 71 infection cleaves a negative regulator for viral internal ribosomal entry site-driven translation. J Virol 2013;87:3828–3838 [CrossRef][PubMed]
    [Google Scholar]
  10. Gao G, Guo X, Goff SP. Inhibition of retroviral RNA production by ZAP, a CCCH-type zinc finger protein. Science 2002;297:1703–1706 [CrossRef][PubMed]
    [Google Scholar]
  11. Kerns JA, Emerman M, Malik HS. Positive selection and increased antiviral activity associated with the PARP-containing isoform of human zinc-finger antiviral protein. PLoS Genet 2008;4:e21 [CrossRef][PubMed]
    [Google Scholar]
  12. Bick MJ, Carroll JW, Gao G, Goff SP, Rice CM et al. Expression of the zinc-finger antiviral protein inhibits alphavirus replication. J Virol 2003;77:11555–11562 [CrossRef][PubMed]
    [Google Scholar]
  13. Mao R, Nie H, Cai D, Zhang J, Liu H et al. Inhibition of hepatitis B virus replication by the host zinc finger antiviral protein. PLoS Pathog 2013;9:e1003494 [CrossRef][PubMed]
    [Google Scholar]
  14. Müller S, Möller P, Bick MJ, Wurr S, Becker S et al. Inhibition of filovirus replication by the zinc finger antiviral protein. J Virol 2007;81:2391–2400 [CrossRef][PubMed]
    [Google Scholar]
  15. Zhu Y, Chen G, Lv F, Wang X, Ji X et al. Zinc-finger antiviral protein inhibits HIV-1 infection by selectively targeting multiply spliced viral mRNAs for degradation. Proc Natl Acad Sci USA 2011;108:15834–15839 [CrossRef][PubMed]
    [Google Scholar]
  16. Li M, Yan K, Wei L, Yang J, Lu C et al. Zinc finger antiviral protein inhibits coxsackievirus B3 virus replication and protects against viral myocarditis. Antiviral Res 2015;123:50–61 [CrossRef][PubMed]
    [Google Scholar]
  17. Guo X, Carroll JW, Macdonald MR, Goff SP, Gao G. The zinc finger antiviral protein directly binds to specific viral mRNAs through the CCCH zinc finger motifs. J Virol 2004;78:12781–12787 [CrossRef][PubMed]
    [Google Scholar]
  18. Guo X, Ma J, Sun J, Gao G. The zinc-finger antiviral protein recruits the RNA processing exosome to degrade the target mRNA. Proc Natl Acad Sci USA 2007;104:151–156 [CrossRef][PubMed]
    [Google Scholar]
  19. Chen G, Guo X, Lv F, Xu Y, Gao G. p72 DEAD box RNA helicase is required for optimal function of the zinc-finger antiviral protein. Proc Natl Acad Sci USA 2008;105:4352–4357 [CrossRef][PubMed]
    [Google Scholar]
  20. Chen S, Xu Y, Zhang K, Wang X, Sun J et al. Structure of N-terminal domain of ZAP indicates how a zinc-finger protein recognizes complex RNA. Nat Struct Mol Biol 2012;19:430–435 [CrossRef][PubMed]
    [Google Scholar]
  21. Xuan Y, Gong D, Qi J, Han C, Deng H et al. ZAP inhibits murine gammaherpesvirus 68 ORF64 expression and is antagonized by RTA. J Virol 2013;87:2735–2743 [CrossRef][PubMed]
    [Google Scholar]
  22. Zhu Y, Wang X, Goff SP, Gao G. Translational repression precedes and is required for ZAP-mediated mRNA decay. Embo J 2012;31:4236–4246 [CrossRef][PubMed]
    [Google Scholar]
  23. Wang J, Fan T, Yao X, Wu Z, Guo L et al. Crystal structures of enterovirus 71 3C protease complexed with rupintrivir reveal the roles of catalytically important residues. J Virol 2011;85:10021–10030 [CrossRef][PubMed]
    [Google Scholar]
  24. Rebsamen M, Meylan E, Curran J, Tschopp J. The antiviral adaptor proteins Cardif and Trif are processed and inactivated by caspases. Cell Death Differ 2008;15:1804–1811 [CrossRef][PubMed]
    [Google Scholar]
  25. Xue Q, Zhou Z, Lei X, Liu X, He B et al. TRIM38 negatively regulates TLR3-mediated IFN-β signaling by targeting TRIF for degradation. PLoS One 2012;7:e46825 [CrossRef][PubMed]
    [Google Scholar]
  26. Li J, Hou N, Faried A, Tsutsumi S, Takeuchi T et al. Inhibition of autophagy by 3-MA enhances the effect of 5-FU-induced apoptosis in colon cancer cells. Ann Surg Oncol 2009;16:761–771 [CrossRef][PubMed]
    [Google Scholar]
  27. Shih SR, Chiang C, Chen TC, Wu CN, Hsu JT et al. Mutations at KFRDI and VGK domains of enterovirus 71 3C protease affect its RNA binding and proteolytic activities. J Biomed Sci 2004;11:239–248 [CrossRef][PubMed]
    [Google Scholar]
  28. Wang X, Tu F, Zhu Y, Gao G. Zinc-finger antiviral protein inhibits XMRV infection. PLoS One 2012;7:e39159 [CrossRef][PubMed]
    [Google Scholar]
  29. Kawai T, Akira S. Innate immune recognition of viral infection. Nat Immunol 2006;7:131–137 [CrossRef][PubMed]
    [Google Scholar]
  30. Schneider WM, Chevillotte MD, Rice CM. Interferon-stimulated genes: a complex web of host defenses. Annu Rev Immunol 2014;32:513–545 [CrossRef][PubMed]
    [Google Scholar]
  31. Wang N, Dong Q, Li J, Jangra RK, Fan M et al. Viral induction of the zinc finger antiviral protein is IRF3-dependent but NF-κB-independent. J Biol Chem 2010;285:6080–6090 [CrossRef][PubMed]
    [Google Scholar]
  32. Chang LY, Huang LM, Gau SS, Wu YY, Hsia SH et al. Neurodevelopment and cognition in children after enterovirus 71 infection. N Engl J Med 2007;356:1226–1234 [CrossRef][PubMed]
    [Google Scholar]
  33. Ho M, Chen ER, Hsu KH, Twu SJ, Chen KT et al. An epidemic of enterovirus 71 infection in Taiwan. Taiwan enterovirus epidemic working group. N Engl J Med 1999;341:929–935 [CrossRef][PubMed]
    [Google Scholar]
  34. Huang CC, Liu CC, Chang YC, Chen CY, Wang ST et al. Neurologic complications in children with enterovirus 71 infection. N Engl J Med 1999;341:936–942 [CrossRef][PubMed]
    [Google Scholar]
  35. Lin YW, Chang KC, Kao CM, Chang SP, Tung YY et al. Lymphocyte and antibody responses reduce enterovirus 71 lethality in mice by decreasing tissue viral loads. J Virol 2009;83:6477–6483 [CrossRef][PubMed]
    [Google Scholar]
  36. Falk MM, Grigera PR, Bergmann IE, Zibert A, Multhaup G et al. Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3. J Virol 1990;64:748–756[PubMed]
    [Google Scholar]
  37. Schlax PE, Zhang J, Lewis E, Planchart A, Lawson TG. Degradation of the encephalomyocarditis virus and hepatitis A virus 3C proteases by the ubiquitin/26S proteasome system in vivo. Virology 2007;360:350–363 [CrossRef][PubMed]
    [Google Scholar]
  38. Lawson TG, Smith LL, Palmenberg AC, Thach RE. Inducible expression of encephalomyocarditis virus 3C protease activity in stably transformed mouse cell lines. J Virol 1989;63:5013–5022[PubMed]
    [Google Scholar]
  39. Zhu Y, Gao G. ZAP-mediated mRNA degradation. RNA Biol 2008;5:65–67 [CrossRef][PubMed]
    [Google Scholar]
  40. Liu CH, Zhou L, Chen G, Krug RM. Battle between influenza A virus and a newly identified antiviral activity of the PARP-containing ZAPL protein. Proc Natl Acad Sci USA 2015;112:14048–14053 [CrossRef][PubMed]
    [Google Scholar]
  41. Aravind L. The WWE domain: a common interaction module in protein ubiquitination and ADP ribosylation. Trends Biochem Sci 2001;26:273–275 [CrossRef][PubMed]
    [Google Scholar]
  42. Charron G, Li MM, MacDonald MR, Hang HC. Prenylome profiling reveals S-farnesylation is crucial for membrane targeting and antiviral activity of ZAP long-isoform. Proc Natl Acad Sci USA 2013;110:11085–11090 [CrossRef][PubMed]
    [Google Scholar]
  43. Gläsker S, Töller M, Kümmerer BM. The alternate triad motif of the poly(ADP-ribose) polymerase-like domain of the human zinc finger antiviral protein is essential for its antiviral activity. J Gen Virol 2014;95:816–822 [CrossRef][PubMed]
    [Google Scholar]
  44. Zheng Z, Ke X, Wang M, He S, Li Q et al. Human microRNA hsa-miR-296-5p suppresses enterovirus 71 replication by targeting the viral genome. J Virol 2013;87:5645–5656 [CrossRef][PubMed]
    [Google Scholar]
  45. Pizzi M. Sampling variation of the fifty percent end-point, determined by the Reed-Muench (Behrens) method. Hum Biol 1950;22:151–190[PubMed]
    [Google Scholar]
  46. Chen Z, Sahashi Y, Matsuo K, Asanuma H, Takahashi H et al. Comparison of the ability of viral protein-expressing plasmid DNAs to protect against influenza. Vaccine 1998;16:1544–1549 [CrossRef][PubMed]
    [Google Scholar]
  47. Deng C, Li X, Liu S, Xu L, Ye H et al. Development and characterization of a clinical strain of Coxsackievirus A16 and an eGFP infectious clone. Virol Sin 2015;30:269–276 [CrossRef][PubMed]
    [Google Scholar]
  48. Zhang Z, Zheng Z, Luo H, Meng J, Li H et al. Human bocavirus NP1 inhibits IFN-β production by blocking association of IFN regulatory factor 3 with IFNB promoter. J Immunol 2012;189:1144–1153 [CrossRef][PubMed]
    [Google Scholar]
  49. Hayakawa S, Shiratori S, Yamato H, Kameyama T, Kitatsuji C et al. ZAPS is a potent stimulator of signaling mediated by the RNA helicase RIG-I during antiviral responses. Nat Immunol 2011;12:37–44 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000982
Loading
/content/journal/jgv/10.1099/jgv.0.000982
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error