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Volume 102, Issue 3

Nuclear localization of Zika virus NS5 contributes to suppression of type I interferon production and response Open Access

Abstract

Zika virus (ZIKV) is an emerging mosquito-borne flavivirus, which caused an unprecedented epidemic in Latin America. Among all viral non-structural proteins in flavivirus, NS5 is the most highly conserved and has multiple crucial functions, including participating in viral RNA replication and suppressing host innate immunity. Although ZIKV NS5 prominently localizes in the nucleus during infection, its specific nuclear localization signal (NLS), and its role in viral replication and pathogenesis remain controversial. Here, we identified aa 11–90 and aa 370–406 regions that contain NLSs, which are critical for nuclear localization of ZIKV NS5. Further experiments demonstrated that nuclear localization of ZIKV NS5 predominantly participates in suppression of interferon regulatory factor 3 (IRF3)-mediated activation of type I IFN (IFN-I) transcription and inhibition of IFN-I downstream response independent of its effect on signal transducers and activators of transcription 2 (STAT2) degradation. These results suggest that subcellular localization of NS5 is important for its function on innate immune suppression, which provides new insight into ZIKV pathogenesis.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): immune evasion , NS5 , nuclear localization signal (NLS) , type I interferon (IFN-I) and Zika virus (ZIKV)
Funding
This study was supported by the:
  • Natural Science Foundation of Hubei Province (Award 2019CFA010)
    • Principle Award Recipient: Shengbo Cao
  • Fundamental Research Funds for the Central Universities (Award 2662018QD025)
    • Principle Award Recipient: Jing Ye
  • National Natural Science Foundation of China (Award 31972721)
    • Principle Award Recipient: Jing Ye
  • National Natural Science Foundation of China (Award 31572517)
    • Principle Award Recipient: Shengbo Cao
  • National Natural Science Foundation of China (Award 31825025)
    • Principle Award Recipient: Shengbo Cao
  • National Key Research and Development Program of China (Award 2016YFD0500407)
    • Principle Award Recipient: Shengbo Cao
© 2021 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License.
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2019-12-20
2024-04-26
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References

  1. Dick GWA, Kitchen SF, Haddow AJ. Zika virus. I. isolations and serological specificity. Trans R Soc Trop Med Hyg 1952; 46:509–520 [View Article][PubMed]
     [Google Scholar]
  2. Faria NR, Azevedo RdSdS, Kraemer MUG, Souza R, Cunha MS et al. Zika virus in the Americas: early epidemiological and genetic findings. Science 2016; 352:345–349 [View Article]
     [Google Scholar]
  3. Mlakar J, Korva M, Tul N, Popović M, Poljšak-Prijatelj M et al. Zika virus associated with microcephaly. N Engl J Med 2016; 374:951–958 [View Article]
     [Google Scholar]
  4. Cao-Lormeau VM, Blake A, Mons S, Lastère S, Roche C et al. Guillain-Barré syndrome outbreak associated with Zika virus infection in French Polynesia: a case-control study. The Lancet 2016; 387:1531–1539 [View Article]
     [Google Scholar]
  5. Chambers TJ, Hahn CS, Galler R, Rice CM. Flavivirus genome organization, expression, and replication. Annu Rev Microbiol 1990; 44:649–688 [View Article]
     [Google Scholar]
  6. Zhu Z, Chan JFW, Tee KM, Choi GKY, Lau SKP et al. Comparative genomic analysis of pre-epidemic and epidemic Zika virus strains for virological factors potentially associated with the rapidly expanding epidemic. Emerg Microbes Infect 2016; 5:e221–12 [View Article][PubMed]
     [Google Scholar]
  7. Zhao B, Yi G, Du F, Chuang YC, Vaughan RC et al. Structure and function of the Zika virus full-length NS5 protein. Nat Commun 2017; 8:14762 [View Article][PubMed]
     [Google Scholar]
  8. Wang B, Tan XF, Thurmond S, Zhang ZM, Lin A et al. The structure of Zika virus NS5 reveals a conserved domain conformation. Nat Commun 2017; 8:14763 [View Article][PubMed]
     [Google Scholar]
  9. Klema V, Padmanabhan R, Choi K. Flaviviral Replication Complex: Coordination between RNA Synthesis and 5’-RNA Capping. Viruses 2015; 7:4640–4656 [View Article]
     [Google Scholar]
  10. Ye J, Chen Z, Zhang B, Miao H, Zohaib A et al. Heat shock protein 70 is associated with replicase complex of Japanese encephalitis virus and positively regulates viral genome replication. PLoS One 2013; 8:e75188 [View Article]
     [Google Scholar]
  11. Chen Z, Ye J, Ashraf U, Li Y, Wei S et al. MicroRNA-33a-5p modulates Japanese encephalitis virus replication by targeting eukaryotic translation elongation factor 1A1. J Virol 2016; 90:3722–3734 [View Article]
     [Google Scholar]
  12. Grant A, Ponia SS, Tripathi S, Balasubramaniam V, Miorin L et al. Zika virus targets human STAT2 to inhibit type I interferon signaling. Cell Host Microbe 2016; 19:882–890 [View Article]
     [Google Scholar]
  13. Pryor MJ, Rawlinson SM, Butcher RE, Barton CL, Waterhouse TA et al. Nuclear localization of dengue virus nonstructural protein 5 through its importin α/β-Recognized nuclear localization sequences is integral to viral infection. Traffic 2007; 8:795–807 [View Article]
     [Google Scholar]
  14. Buckley A, Gaidamovich S, Turchinskaya A, Gould EA. Monoclonal antibodies identify the NS5 yellow fever virus non-structural protein in the nuclei of infected cells. J Gen Virol 1992; 73:1125–1130 [View Article][PubMed]
     [Google Scholar]
  15. Rawlinson SM, Pryor MJ, Wright PJ, Jans DA. Crm1-Mediated nuclear export of dengue virus RNA polymerase NS5 modulates interleukin-8 induction and virus production. J Biol Chem 2009; 284:15589–15597 [View Article][PubMed]
     [Google Scholar]
  16. De Maio FA, Risso G, Iglesias NG, Shah P, Pozzi B et al. The dengue virus NS5 protein Intrudes in the cellular spliceosome and modulates splicing. PLoS Pathog 2016; 12:e1005841 [View Article]
     [Google Scholar]
  17. Lange A, Mills RE, Lange CJ, Stewart M, Devine SE et al. Classical nuclear localization signals: definition, function, and interaction with importin alpha. J Biol Chem 2007; 282:5101–5105 [View Article][PubMed]
     [Google Scholar]
  18. Forwood JK, Brooks A, Briggs LJ, Xiao CY, Jans DA et al. The 37-amino-acid interdomain of dengue virus NS5 protein contains a functional NLS and inhibitory CK2 site. Biochem Biophys Res Commun 1999; 257:731–737 [View Article][PubMed]
     [Google Scholar]
  19. Versteeg GA, García-Sastre A. Viral tricks to grid-lock the type I interferon system. Curr Opin Microbiol 2010; 13:508–516 [View Article]
     [Google Scholar]
  20. Ye J, Chen Z, Li Y, Zhao Z, He W et al. Japanese encephalitis virus NS5 inhibits type I interferon (IFN) production by blocking the nuclear translocation of IFN regulatory factor 3 and NF-κB. J Virol 2017; 91: [View Article]
     [Google Scholar]
  21. Lubick KJ, Robertson SJ, McNally KL, Freedman BA, Rasmussen AL et al. Flavivirus antagonism of type I interferon signaling reveals prolidase as a regulator of IFNAR1 surface expression. Cell Host Microbe 2015; 18:61–74 [View Article]
     [Google Scholar]
  22. Kumar A, Hou S, Airo AM, Limonta D, Mancinelli V et al. Zika virus inhibits type‐I interferon production and downstream signaling. EMBO Rep 2016; 17:1766–1775 [View Article]
     [Google Scholar]
  23. Hertzog J, Dias Junior AG, Rigby RE, Donald CL, Mayer A et al. Infection with a Brazilian isolate of Zika virus generates RIG‐I stimulatory RNA and the viral NS5 protein blocks type I IFN induction and signaling. Eur J Immunol 2018; 48:1120–1136 [View Article]
     [Google Scholar]
  24. Maertens G, Cherepanov P, Debyser Z, Engelborghs Y, Engelman A. Identification and characterization of a functional nuclear localization signal in the HIV-1 integrase interactor LEDGF/p75. Journal of Biological Chemistry 2004; 279:33421–33429 [View Article]
     [Google Scholar]
  25. Woodward CL, Wang Y, Dixon WJ, Htun H, Chow SA. Subcellular localization of feline immunodeficiency virus integrase and mapping of its karyophilic determinant. J Virol 2003; 77:4516–4527 [View Article]
     [Google Scholar]
  26. Kalderon D, Roberts BL, Richardson WD, Smith AE. A short amino acid sequence able to specify nuclear location. Cell 1984; 39:499–509 [View Article]
     [Google Scholar]
  27. Kosugi S, Hasebe M, Matsumura N, Takashima H, Miyamoto-Sato E et al. Six classes of nuclear localization signals specific to different binding grooves of importin alpha. J Biol Chem 2009; 284:478–485 [View Article][PubMed]
     [Google Scholar]
  28. Wang C, Yang SNY, Smith K, Forwood JK, Jans DA. Nuclear import inhibitor N -(4-hydroxyphenyl) retinamide targets Zika virus (ZIKV) nonstructural protein 5 to inhibit ZIKV infection. Biochem Biophys Res Commun 2017; 493:1555–1559 [View Article]
     [Google Scholar]
  29. Pichlmair A. Reis E Sousa C. innate recognition of viruses. Immunity 2007; 27:370–383
     [Google Scholar]
  30. Xia H, Luo H, Shan C, Muruato AE, Nunes BTD et al. An evolutionary NS1 mutation enhances Zika virus evasion of host interferon induction. Nat Commun 2018; 9:414 [View Article]
     [Google Scholar]
  31. Morrison J, Laurent-Rolle M, Maestre AM, Rajsbaum R, Pisanelli G et al. Dengue virus co-opts UBR4 to degrade STAT2 and antagonize type I interferon signaling. PLoS Pathog 2013; 9:e1003265 [View Article]
     [Google Scholar]
  32. Fontes MRM, Teh T, Jans D, Brinkworth RI, Kobe B. Structural basis for the specificity of bipartite nuclear localization sequence binding by importin-alpha. J Biol Chem 2003; 278:27981–27987 [View Article][PubMed]
     [Google Scholar]
  33. Brooks AJ, Johansson M, John AV, Xu Y, Jans DA et al. The interdomain region of dengue NS5 protein that binds to the viral helicase NS3 contains independently functional importin beta 1 and importin alpha/beta-recognized nuclear localization signals. J Biol Chem 2002; 277:36399–36407 [View Article][PubMed]
     [Google Scholar]
  34. Lopez-Denman AJ, Russo A, Wagstaff KM, White PA, Jans DA et al. Nucleocytoplasmic shuttling of the West Nile virus RNA-dependent RNA polymerase NS5 is critical to infection. Cell Microbiol 2018; 20:e12848 [View Article]
     [Google Scholar]
  35. Tay MYF, Smith K, Ng IHW, Chan KWK, Zhao Y et al. The C-terminal 18 amino acid region of dengue virus NS5 regulates its subcellular localization and contains a conserved arginine residue essential for infectious virus production. PLoS Pathog 2016; 12:e1005886
     [Google Scholar]
  36. Kosugi S, Hasebe M, Tomita M, Yanagawa H. Systematic identification of cell cycle-dependent yeast nucleocytoplasmic shuttling proteins by prediction of composite motifs. Proc Natl Acad Sci USA 2009; 106:10171–10176 [View Article][PubMed]
     [Google Scholar]
  37. Bowen JR, Quicke KM, Maddur MS, O’Neal JT, McDonald CE et al. Zika virus antagonizes type I interferon responses during infection of human dendritic cells. PLoS Pathog 2017; 13:e1006164 [View Article]
     [Google Scholar]
  38. Ivashkiv LB, Donlin LT. Regulation of type I interferon responses. Nat Rev Immunol 2014; 14:36–49 [View Article]
     [Google Scholar]
  39. LJ H, Hung LF, Weng CY, WL W, Chou P et al. Dengue virus type 2 antagonizes IFN-alpha but not IFN-gamma antiviral effect via down-regulating Tyk2-STAT signaling in the human dendritic cell. J Immuno 2005; 174:8163–8172
     [Google Scholar]
  40. Guo JT, Hayashi J, Seeger C. West Nile virus inhibits the signal transduction pathway of alpha interferon. J Virol 2005; 79:1343–1350 [View Article][PubMed]
     [Google Scholar]
  41. Mukherjee A, Di Bisceglie AM, Ray RB. Hepatitis C virus-mediated enhancement of microRNA miR-373 impairs the JAK/STAT signaling pathway. J Virol 2015; 89:3356–3365 [View Article]
     [Google Scholar]
  42. Zhang R, Fang L, Wang D, Cai K, Zhang H et al. Porcine bocavirus NP1 negatively regulates interferon signaling pathway by targeting the DNA-binding domain of IRF9. Virology 2015; 485:414–421 [View Article]
     [Google Scholar]
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Nuclear localization of Zika virus NS5 contributes to suppression of type I interferon production and response
J. Gen. Virol. 102, 001376 (2021); https://doi.org/10.1099/jgv.0.001376
/content/journal/jgv/10.1099/jgv.0.001376
/content/journal/jgv/10.1099/jgv.0.001376
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