1887

Abstract

The accessory protein, PB1-F2, of influenza A virus (IAV) functions in a chicken host to prolong infectious virus shedding and thus the transmission window. Here we show that this delay in virus clearance by PB1-F2 in chickens is accompanied by reduced transcript levels of type 1 interferon (IFN)-induced genes and NFκB-activated pro-inflammation cytokines. In vitro, two avian influenza isolate-derived PB1-F2 proteins, H9N2 UDL01 and H5N1 5092, exhibited the same antagonism of the IFN and pro-inflammation induction pathways seen in vivo, but to different extents. The two PB1-F2 proteins had different cellular localization in chicken cells, with H5N1 5092 being predominantly mitochondrial-associated and H9N2 UDL being cytoplasmic but not mitochondrial-localized. We hypothesized that PB1-F2 localization might influence the functionality of the protein during infection and that the protein sequence could alter cellular localization. We demonstrated that the sequence of the C-terminus of PB1-F2 determined cytoplasmic localization in chicken cells and this was linked with protein instability. Mitochondrial localization of PB1-F2 resulted in reduced antagonism of an NFκB-dependent promoter. In parallel, mitochondrial localization of PB1-F2 increased the potency of chicken IFN 2 induction antagonism. We suggest that mitochondrial localization of PB1-F2 restricts interaction with cytoplasmic-located IKKβ, reducing NFκB-responsive promoter antagonism, but enhances antagonism of the IFN2 promoter through interaction with the mitochondrial adaptor MAVS. Our study highlights the differential mechanisms by which IAV PB1-F2 protein can dampen the avian host innate signalling response.

Keyword(s): chicken , IKKβ , influenza virus , MAVS and PB1-F2
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001220
2019-01-23
2021-10-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/100/3/414.html?itemId=/content/journal/jgv/10.1099/jgv.0.001220&mimeType=html&fmt=ahah

References

  1. Basuno E, Yusdja Y, Ilham N. Socio-economic impacts of avian influenza outbreaks on small-scale producers in Indonesia. Transbound Emerg Dis 2010; 57:7–10 [View Article][PubMed]
    [Google Scholar]
  2. Govindaraj G, Sridevi R, Nandakumar SN, Vineet R, Rajeev P et al. Economic impacts of avian influenza outbreaks in Kerala, India. Transbound Emerg Dis 2018; 65:e361e372 [View Article][PubMed]
    [Google Scholar]
  3. Qi X, Jiang D, Wang H, Zhuang D, Ma J et al. Calculating the burden of disease of avian-origin H7N9 infections in China. BMJ Open 2014; 4:e004189 [View Article][PubMed]
    [Google Scholar]
  4. Vasin AV, Temkina OA, Egorov VV, Klotchenko SA, Plotnikova MA et al. Molecular mechanisms enhancing the proteome of influenza A viruses: an overview of recently discovered proteins. Virus Res 2014; 185:53–63 [View Article][PubMed]
    [Google Scholar]
  5. Chen W, Calvo PA, Malide D, Gibbs J, Schubert U et al. A novel influenza A virus mitochondrial protein that induces cell death. Nat Med 2001; 7:1306–1312 [View Article][PubMed]
    [Google Scholar]
  6. James J, Howard W, Iqbal M, Nair VK, Barclay WS et al. Influenza A virus PB1-F2 protein prolongs viral shedding in chickens lengthening the transmission window. J Gen Virol 2016; 97:2516–2527 [View Article][PubMed]
    [Google Scholar]
  7. Zell R, Krumbholz A, Eitner A, Krieg R, Halbhuber KJ et al. Prevalence of PB1-F2 of influenza A viruses. J Gen Virol 2007; 88:536–546 [View Article][PubMed]
    [Google Scholar]
  8. Kamal R, Alymova I, York I. Evolution and Virulence of Influenza A Virus Protein PB1-F2. Int J Mol Sci 2017; 19:96 [View Article]
    [Google Scholar]
  9. Klemm C, Boergeling Y, Ludwig S, Ehrhardt C. Immunomodulatory Nonstructural Proteins of Influenza A Viruses. Trends Microbiol 2018; 26:624–636 [View Article][PubMed]
    [Google Scholar]
  10. Košík I, Práznovská M, Košíková M, Bobišová Z, Hollý J et al. The ubiquitination of the influenza A virus PB1-F2 protein is crucial for its biological function. PLoS One 2015; 10:e0118477 [View Article][PubMed]
    [Google Scholar]
  11. Leymarie O, Embury-Hyatt C, Chevalier C, Jouneau L, Moroldo M et al. PB1-F2 attenuates virulence of highly pathogenic avian H5N1 influenza virus in chickens. PLoS One 2014; 9:e100679 [View Article][PubMed]
    [Google Scholar]
  12. de Jong MD, Simmons CP, Thanh TT, Hien VM, Smith GJ et al. Fatal outcome of human influenza A (H5N1) is associated with high viral load and hypercytokinemia. Nat Med 2006; 12:1203–1207 [View Article][PubMed]
    [Google Scholar]
  13. Abdel-Ghafar AN, Chotpitayasunondh T, Gao Z, Hayden FG, Nguyen DH et al. Update on avian influenza A (H5N1) virus infection in humans. N Engl J Med 2008; 358:261–273 [View Article][PubMed]
    [Google Scholar]
  14. Yoshizumi T, Ichinohe T, Sasaki O, Otera H, Kawabata S et al. Influenza A virus protein PB1-F2 translocates into mitochondria via Tom40 channels and impairs innate immunity. Nat Commun 2014; 5:4713 [View Article][PubMed]
    [Google Scholar]
  15. Chang P, Kuchipudi SV, Mellits KH, Sebastian S, James J et al. Early apoptosis of porcine alveolar macrophages limits avian influenza virus replication and pro-inflammatory dysregulation. Sci Rep 2015; 5:17999 [View Article][PubMed]
    [Google Scholar]
  16. Dudek SE, Wixler L, Nordhoff C, Nordmann A, Anhlan D et al. The influenza virus PB1-F2 protein has interferon antagonistic activity. Biol Chem 2011; 392:1135–1144 [View Article][PubMed]
    [Google Scholar]
  17. Leymarie O, Meyer L, Tafforeau L, Lotteau V, Costa BD et al. Influenza virus protein PB1-F2 interacts with CALCOCO2 (NDP52) to modulate innate immune response. J Gen Virol 2017; 98:1196–1208 [View Article][PubMed]
    [Google Scholar]
  18. Conenello GM, Tisoncik JR, Rosenzweig E, Varga ZT, Palese P et al. A single N66S mutation in the PB1-F2 protein of influenza A virus increases virulence by inhibiting the early interferon response in vivo. J Virol 2011; 85:652–662 [View Article][PubMed]
    [Google Scholar]
  19. Varga ZT, Grant A, Manicassamy B, Palese P. Influenza virus protein PB1-F2 inhibits the induction of type I interferon by binding to MAVS and decreasing mitochondrial membrane potential. J Virol 2012; 86:8359–8366 [View Article][PubMed]
    [Google Scholar]
  20. Varga ZT, Ramos I, Hai R, Schmolke M, García-Sastre A et al. The influenza virus protein PB1-F2 inhibits the induction of type I interferon at the level of the MAVS adaptor protein. PLoS Pathog 2011; 7:e1002067 [View Article][PubMed]
    [Google Scholar]
  21. Chen S, Cheng A, Wang M. Innate sensing of viruses by pattern recognition receptors in birds. Vet Res 2013; 44:82 [View Article][PubMed]
    [Google Scholar]
  22. Cheng YY, Yang SR, Wang YT, Lin YH, Chen CJ. Amino acid residues 68-71 contribute to influenza A virus PB1-F2 protein stability and functions. Front Microbiol 2017; 8:692 [View Article][PubMed]
    [Google Scholar]
  23. Chen CJ, Chen GW, Wang CH, Huang CH, Wang YC et al. Differential localization and function of PB1-F2 derived from different strains of influenza A virus. J Virol 2010; 84:10051–10062 [View Article][PubMed]
    [Google Scholar]
  24. Gibbs JS, Malide D, Hornung F, Bennink JR, Yewdell JW. The influenza A virus PB1-F2 protein targets the inner mitochondrial membrane via a predicted basic amphipathic helix that disrupts mitochondrial function. J Virol 2003; 77:7214–7224 [View Article][PubMed]
    [Google Scholar]
  25. Yamada H, Chounan R, Higashi Y, Kurihara N, Kido H. Mitochondrial targeting sequence of the influenza A virus PB1-F2 protein and its function in mitochondria. FEBS Lett 2004; 578:331–336 [View Article][PubMed]
    [Google Scholar]
  26. Reuter A, Soubies S, Härtle S, Schusser B, Kaspers B et al. Antiviral activity of lambda interferon in chickens. J Virol 2014; 88:2835–2843 [View Article][PubMed]
    [Google Scholar]
  27. Liniger M, Summerfield A, Zimmer G, Mccullough KC, Ruggli N. Chicken cells sense influenza A virus infection through MDA5 and CARDIF signaling involving LGP2. J Virol 2012; 86:705–717 [View Article][PubMed]
    [Google Scholar]
  28. Reis AL, Mccauley JW. The influenza virus protein PB1-F2 interacts with IKKβ and modulates NF-κB signalling. PLoS One 2013; 8:e63852 [View Article][PubMed]
    [Google Scholar]
  29. Miodek A, Sauriat-Dorizon H, Chevalier C, Delmas B, Vidic J et al. Direct electrochemical detection of PB1-F2 protein of influenza A virus in infected cells. Biosens Bioelectron 2014; 59:6–13 [View Article][PubMed]
    [Google Scholar]
  30. Miodek A, Vidic J, Sauriat-Dorizon H, Richard CA, Le Goffic R et al. Electrochemical detection of the oligomerization of PB1-F2 influenza A virus protein in infected cells. Anal Chem 2014; 86:9098–9105 [View Article][PubMed]
    [Google Scholar]
  31. Wei P, Li W, Zi H, Cunningham M, Guo Y et al. Epidemiological and molecular characteristics of the PB1-F2 proteins in H7N9 influenza viruses, Jiangsu. Biomed Res Int 2015; 2015:1–8 [View Article][PubMed]
    [Google Scholar]
  32. Holmes EC, Lipman DJ, Zamarin D, Yewdell JW. Comment on "Large-scale sequence analysis of avian influenza isolates". Science 2006; 313:1573b [View Article][PubMed]
    [Google Scholar]
  33. Marais R, Light Y, Paterson HF, Marshall CJ. Ras recruits Raf-1 to the plasma membrane for activation by tyrosine phosphorylation. EMBO J 1995; 14:3136–3145 [View Article][PubMed]
    [Google Scholar]
  34. Horner SM, Liu HM, Park HS, Briley J, Gale M. Mitochondrial-associated endoplasmic reticulum membranes (MAM) form innate immune synapses and are targeted by hepatitis C virus. Proc Natl Acad Sci USA 2011; 108:14590–14595 [View Article][PubMed]
    [Google Scholar]
  35. Carpenter AE, Jones TR, Lamprecht MR, Clarke C, Kang IH et al. CellProfiler: image analysis software for identifying and quantifying cell phenotypes. Genome Biol 2006; 7:R100 [View Article][PubMed]
    [Google Scholar]
  36. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001; 29:45e–45 [View Article][PubMed]
    [Google Scholar]
  37. Spackman E, Senne DA, Myers TJ, Bulaga LL, Garber LP et al. Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. J Clin Microbiol 2002; 40:3256–3260 [View Article][PubMed]
    [Google Scholar]
  38. Berger Rentsch M, Zimmer G. A vesicular stomatitis virus replicon-based bioassay for the rapid and sensitive determination of multi-species type I interferon. PLoS One 2011; 6:e25858 [View Article][PubMed]
    [Google Scholar]
  39. Moncorgé O, Long JS, Cauldwell AV, Zhou H, Lycett SJ et al. Investigation of influenza virus polymerase activity in pig cells. J Virol 2013; 87:384–394 [View Article][PubMed]
    [Google Scholar]
  40. Giotis ES, Robey RC, Skinner NG, Tomlinson CD, Goodbourn S et al. Chicken interferome: avian interferon-stimulated genes identified by microarray and RNA-seq of primary chick embryo fibroblasts treated with a chicken type I interferon (IFN-α). Vet Res 2016; 47:75 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001220
Loading
/content/journal/jgv/10.1099/jgv.0.001220
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Most cited this month Most Cited RSS feed

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