1887

Abstract

Post-translational modification plays a critical role in viral replication. Previously we reported that neddylation of PB2 of influenza A virus (IAV) can inhibit viral replication. However, we found that NEDD8 overexpression can still inhibit the replication of PB2 K699R mutant viruses, implying that other viral protein(s) can be neddylated. In this study, we revealed that M1 of IAV can also be modified by NEDD8. We found that the E3 ligase HDM2 significantly promotes M1 neddylation. Furthermore, we identified M1 K187 as the major neddylation site. We generated an IAV M1 K187R mutant (WSN-M1 K187R) and compared the growth of wild-type and mutant viruses in Madin–Darby canine kidney (MDCK) cells. The data showed that the replication of WSN-M1 K187R was more efficient than that of wild-type WSN. More importantly, we observed that overexpression of NEDD8 inhibited the replication of the wild-type WSN more effectively than that of WSN-M1 K187R. In addition, we found that the neddylation-deficient M1 mutant (M1 K187R) had a longer half-life than that of wild-type M1, indicating that the neddylation of M1 reduces stability. Then we performed a viral infection assay and found that WSN-M1 K187R exhibited greater virulence in mice than wild-type WSN, suggesting that the neddylation of M1 reduced IAV replication . In conclusion, we uncovered that neddylation of M1 by HDM2 negatively regulates the stability of M1, which in turn inhibits viral replication.

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2020-10-05
2024-04-25
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References

  1. Lamb RA, Choppin PW. The gene structure and replication of influenza virus. Annu Rev Biochem 1983; 52:467–506 [View Article][PubMed]
    [Google Scholar]
  2. Resa-Infante P, Jorba N, Coloma R, Ortin J. The influenza virus RNA synthesis machine: advances in its structure and function. RNA Biol 2011; 8:207–215 [View Article][PubMed]
    [Google Scholar]
  3. Arranz R, Coloma R, Chichón FJ, Conesa JJ, Carrascosa JL et al. The structure of native influenza virion ribonucleoproteins. Science 2012; 338:1634–1637 [View Article][PubMed]
    [Google Scholar]
  4. Li KS, Guan Y, Wang J, Smith GJD, Xu KM et al. Genesis of a highly pathogenic and potentially pandemic H5N1 influenza virus in eastern Asia. Nature 2004; 430:209–213 [View Article][PubMed]
    [Google Scholar]
  5. Boyd M, Clezy K, Lindley R, Pearce R. Pandemic influenza: clinical issues. Med J Aust 2006; 185:S44–47 [View Article][PubMed]
    [Google Scholar]
  6. Seo SH, Hoffmann E, Webster RG. Lethal H5N1 influenza viruses escape host anti-viral cytokine responses. Nat Med 2002; 8:950–954 [View Article][PubMed]
    [Google Scholar]
  7. Zheng W, Li J, Wang S, Cao S, Jiang J et al. Phosphorylation controls the nuclear-cytoplasmic shuttling of influenza A virus nucleoprotein. J Virol 2015; 89:5822–5834 [View Article][PubMed]
    [Google Scholar]
  8. Liao TL, Wu CY, Su WC, Jeng KS, Lai MMC. Ubiquitination and deubiquitination of NP protein regulates influenza A virus RNA replication. Embo J 2010; 29:3879–3890 [View Article][PubMed]
    [Google Scholar]
  9. Kirui J, Mondal A, Mehle A. Ubiquitination upregulates influenza virus polymerase function. J Virol 2016; 90:10906–10914 [View Article][PubMed]
    [Google Scholar]
  10. Santos A, Pal S, Chacón J, Meraz K, Gonzalez J et al. SUMOylation affects the interferon blocking activity of the influenza A nonstructural protein NS1 without affecting its stability or cellular localization. J Virol 2013; 87:5602–5620 [View Article][PubMed]
    [Google Scholar]
  11. Zhang T, Ye Z, Yang X, Qin Y, Hu Y et al. Neddylation of PB2 reduces its stability and blocks the replication of influenza A virus. Sci Rep 2017; 7:43691 [View Article][PubMed]
    [Google Scholar]
  12. Whitby FG, Xia G, Pickart CM, Hill CP. Crystal structure of the human ubiquitin-like protein NEDD8 and interactions with ubiquitin pathway enzymes. J Biol Chem 1998; 273:34983–34991 [View Article][PubMed]
    [Google Scholar]
  13. Liakopoulos D, Doenges G, Matuschewski K, Jentsch S. A novel protein modification pathway related to the ubiquitin system. EMBO J 1998; 17:2208–2214 [View Article][PubMed]
    [Google Scholar]
  14. Osaka F, Kawasaki H, Aida N, Saeki M, Chiba T et al. A new NEDD8-ligating system for cullin-4A. Genes Dev 1998; 12:2263–2268 [View Article][PubMed]
    [Google Scholar]
  15. Walden H, Podgorski MS, Schulman BA. Insights into the ubiquitin transfer cascade from the structure of the activating enzyme for NEDD8. Nature 2003; 422:330–334 [View Article][PubMed]
    [Google Scholar]
  16. Wang Z, Zhu WG, Xu X. Ubiquitin-like modifications in the DNA damage response. Mutat Res 2017; 803-805:56–75 [View Article][PubMed]
    [Google Scholar]
  17. Hughes DJ, Wood JJ, Jackson BR, Baquero-Pérez B, Whitehouse A. NEDDylation is essential for Kaposi's sarcoma-associated herpesvirus latency and lytic reactivation and represents a novel anti-KSHV target. PLoS Pathog 2015; 11:e1004771 [View Article][PubMed]
    [Google Scholar]
  18. Liu N, Zhang J, Yang X, Jiao T, Zhao X et al. HDM2 promotes neddylation of hepatitis B virus HBx to enhance its stability and function. J Virol 2017; 91: [View Article][PubMed]
    [Google Scholar]
  19. Nekorchuk MD, Sharifi HJ, Furuya AKM, Jellinger R, de Noronha CMC. HIV relies on neddylation for ubiquitin ligase-mediated functions. Retrovirology 2013; 10:138 [View Article][PubMed]
    [Google Scholar]
  20. Wu CY, Jeng KS, Lai MMC. The sumoylation of matrix protein M1 modulates the assembly and morphogenesis of influenza A virus. J Virol 2011; 85:6618–6628 [View Article][PubMed]
    [Google Scholar]
  21. Su WC, Chen Y-C, Tseng CH, Hsu PW, Tung KF et al. Pooled RNAi screen identifies ubiquitin ligase itch as crucial for influenza A virus release from the endosome during virus entry. Proc Natl Acad Sci U S A 2013; 110:17516–17521 [View Article][PubMed]
    [Google Scholar]
  22. Reinhardt J, Wolff T. The influenza A virus M1 protein interacts with the cellular receptor of activated C kinase (RACK) 1 and can be phosphorylated by protein kinase C. Vet Microbiol 2000; 74:87–100 [View Article][PubMed]
    [Google Scholar]
  23. Squires RB, Noronha J, Hunt V, García-Sastre A, Macken C et al. Influenza research database: an integrated bioinformatics resource for influenza research and surveillance. Influenza Other Respir Viruses 2012; 6:404–416 [View Article][PubMed]
    [Google Scholar]
  24. Liu X, Sun L, Yu M, Wang Z, Xu C et al. Cyclophilin A interacts with influenza A virus M1 protein and impairs the early stage of the viral replication. Cell Microbiol 2009; 11:730–741 [View Article][PubMed]
    [Google Scholar]
  25. Mahesutihan M, Zheng W, Cui L, Li Y, Jiao P et al. CypA regulates AIP4-Mediated M1 ubiquitination of influenza A virus. Virol Sin 2018; 33:440–448 [View Article][PubMed]
    [Google Scholar]
  26. 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 U S A 2015; 112:14048–14053 [View Article][PubMed]
    [Google Scholar]
  27. Di Pietro A, Kajaste-Rudnitski A, Oteiza A, Nicora L, Towers GJ et al. Trim22 inhibits influenza A virus infection by targeting the viral nucleoprotein for degradation. J Virol 2013; 87:4523–4533 [View Article][PubMed]
    [Google Scholar]
  28. Fu B, Wang L, Ding H, Schwamborn JC, Li S et al. Trim32 senses and restricts influenza A virus by ubiquitination of PB1 polymerase. PLoS Pathog 2015; 11:e1004960 [View Article][PubMed]
    [Google Scholar]
  29. Xia C, Vijayan M, Pritzl CJ, Fuchs SY, McDermott AB et al. Hemagglutinin of influenza A virus antagonizes type I interferon (IFN) responses by inducing degradation of type I IFN receptor 1. J Virol 2015; 90:2403–2417 [View Article][PubMed]
    [Google Scholar]
  30. Gack MU, Albrecht RA, Urano T, Inn KS, Huang IC et al. Influenza A virus NS1 targets the ubiquitin ligase TRIM25 to evade recognition by the host viral RNA sensor RIG-I. Cell Host Microbe 2009; 5:439–449 [View Article][PubMed]
    [Google Scholar]
  31. Pizzorno A, Dubois J, Machado D, Cartet G, Traversier A et al. Influenza A viruses alter the stability and antiviral contribution of host E3-ubiquitin ligase Mdm2 during the time-course of infection. Sci Rep 2018; 8:3746 [View Article][PubMed]
    [Google Scholar]
  32. Wang X, Deng X, Yan W, Zhu Z, Shen Y et al. Stabilization of p53 in influenza A virus-infected cells is associated with compromised MDM2-mediated ubiquitination of p53. J Biol Chem 2012; 287:18366–18375 [View Article][PubMed]
    [Google Scholar]
  33. Turrell L, Hutchinson EC, Vreede FT, Fodor E. Regulation of influenza A virus nucleoprotein oligomerization by phosphorylation. J Virol 2015; 89:1452–1455 [View Article][PubMed]
    [Google Scholar]
  34. Wang S, Zhao Z, Bi Y, Sun L, Liu X et al. Tyrosine 132 phosphorylation of influenza A virus M1 protein is crucial for virus replication by controlling the nuclear import of M1. J Virol 2013; 87:6182–6191 [View Article][PubMed]
    [Google Scholar]
  35. Gao S, Wu J, Liu RY, Li J, Song L et al. Interaction of NS2 with AIMP2 facilitates the switch from ubiquitination to sumoylation of M1 in influenza A virus-infected cells. J Virol 2015; 89:300–311 [View Article][PubMed]
    [Google Scholar]
  36. Craig AL, Chrystal JA, Fraser JA, Sphyris N, Lin Y et al. The MDM2 ubiquitination signal in the DNA-binding domain of p53 forms a docking site for calcium calmodulin kinase superfamily members. Mol Cell Biol 2007; 27:3542–3555 [View Article][PubMed]
    [Google Scholar]
  37. Fraser JA, Vojtesek B, Hupp TR. A novel p53 phosphorylation site within the MDM2 ubiquitination signal: I. phosphorylation at SER269 in vivo is linked to inactivation of p53 function. J Biol Chem 2010; 285:37762–37772 [View Article][PubMed]
    [Google Scholar]
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