1887

Abstract

The NS1 protein of influenza virus counters host antiviral defences primarily by antagonizing the type I interferon (IFN) response. Both the N-terminal dsRNA-binding domain and the C-terminal effector domain are required for optimal suppression of host responses during infection. To better understand the regulatory role of the NS1 effector domain, we used an NS1-truncated mutant virus derived from human H1N1 influenza isolate A/Texas/36/91 (Tx/91) and assessed global transcriptional profiles from two independent human lung cell-culture models. Relative to the wild-type Tx/91-induced gene expression, the NS1 mutant virus induced enhanced expression of innate immune genes, specifically NF-κB signalling-pathway genes and IFN-α and -β target genes. We queried an experimentally derived IFN gene set to gauge the proportion of IFN-responsive genes that are suppressed specifically by NS1. We show that the C-terminally truncated NS1 mutant virus is less efficient at suppressing IFN-regulated gene expression associated with activation of antigen-presentation and immune-proteasome pathways. This is the first report integrating genomic analysis from two independent human culture systems, including primary lung cells, using genetically similar H1N1 influenza viruses that differ only in the length of the NS1 protein.

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2011-09-01
2024-03-29
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References

  1. Baskin C. R., Bielefeldt-Ohmann H., García-Sastre A., Tumpey T. M., Van Hoeven N., Carter V. S., Thomas M. J., Proll S., Solórzano A. et al. 2007; Functional genomic and serological analysis of the protective immune response resulting from vaccination of macaques with an NS1-truncated influenza virus. J Virol 81:11817–11827 [View Article][PubMed]
    [Google Scholar]
  2. Billharz R., Zeng H., Proll S. C., Korth M. J., Lederer S., Albrecht R., Goodman A. G., Rosenzweig E., Tumpey T. M. et al. 2009; The NS1 protein of the 1918 pandemic influenza virus blocks host interferon and lipid metabolism pathways. J Virol 83:10557–10570 [View Article][PubMed]
    [Google Scholar]
  3. Brazma A., Hingamp P., Quackenbush J., Sherlock G., Spellman P., Stoeckert C., Aach J., Ansorge W., Ball C. A. et al. 2001; Minimum information about a microarray experiment (MIAME) – toward standards for microarray data. Nat Genet 29:365–371 [View Article][PubMed]
    [Google Scholar]
  4. Chatterjee-Kishore M., Kishore R., Hicklin D. J., Marincola F. M., Ferrone S. 1998; Different requirements for signal transducer and activator of transcription 1alpha and interferon regulatory factor 1 in the regulation of low molecular mass polypeptide 2 and transporter associated with antigen processing 1 gene expression. J Biol Chem 273:16177–16183 [View Article][PubMed]
    [Google Scholar]
  5. Chatterjee-Kishore M., van Den Akker F., Stark G. R. 2000; Adenovirus E1A down-regulates LMP2 transcription by interfering with the binding of stat1 to IRF1. J Biol Chem 275:20406–20411 [View Article][PubMed]
    [Google Scholar]
  6. Chen Z., Li Y., Krug R. M. 1999; Influenza A virus NS1 protein targets poly(A)-binding protein II of the cellular 3′-end processing machinery. EMBO J 18:2273–2283 [View Article][PubMed]
    [Google Scholar]
  7. Chien C. Y., Tejero R., Huang Y., Zimmerman D. E., Ríos C. B., Krug R. M., Montelione G. T. 1997; A novel RNA-binding motif in influenza A virus non-structural protein 1. Nat Struct Biol 4:891–895 [View Article][PubMed]
    [Google Scholar]
  8. Chien C. Y., Xu Y., Xiao R., Aramini J. M., Sahasrabudhe P. V., Krug R. M., Montelione G. T. 2004; Biophysical characterization of the complex between double-stranded RNA and the N-terminal domain of the NS1 protein from influenza A virus: evidence for a novel RNA-binding mode. Biochemistry 43:1950–1962 [View Article][PubMed]
    [Google Scholar]
  9. Darnell J. E. Jr, Kerr I. M., Stark G. R. 1994; Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science 264:1415–1421 [View Article][PubMed]
    [Google Scholar]
  10. de Graaf N., van Helden M. J., Textoris-Taube K., Chiba T., Topham D. J., Kloetzel P. M., Zaiss D. M., Sijts A. J. 2011; PA28 and the proteasome immunosubunits play a central and independent role in the production of MHC class I-binding peptides in vivo . Eur J Immunol 41:926–935 [View Article][PubMed]
    [Google Scholar]
  11. Fernandez-Sesma A., Marukian S., Ebersole B. J., Kaminski D., Park M. S., Yuen T., Sealfon S. C., García-Sastre A., Moran T. M. 2006; Influenza virus evades innate and adaptive immunity via the NS1 protein. J Virol 80:6295–6304 [View Article][PubMed]
    [Google Scholar]
  12. Frontini M., Vijayakumar M., Garvin A., Clarke N. 2009; A ChIP-chip approach reveals a novel role for transcription factor IRF1 in the DNA damage response. Nucleic Acids Res 37:1073–1085 [View Article][PubMed]
    [Google Scholar]
  13. Gack M. U., Albrecht R. A., Urano T., Inn K. S., Huang I. C., Carnero E., Farzan M., Inoue S., Jung J. U., García-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 [View Article][PubMed]
    [Google Scholar]
  14. García-Sastre A. 2001; Inhibition of interferon-mediated antiviral responses by influenza A viruses and other negative-strand RNA viruses. Virology 279:375–384 [View Article][PubMed]
    [Google Scholar]
  15. Garfinkel M. S., Katze M. G. 1992; Translational control by influenza virus. Selective and cap-dependent translation of viral mRNAs in infected cells. J Biol Chem 267:9383–9390[PubMed]
    [Google Scholar]
  16. Geiss G. K., Salvatore M., Tumpey T. M., Carter V. S., Wang X., Basler C. F., Taubenberger J. K., Bumgarner R. E., Palese P. et al. 2002; Cellular transcriptional profiling in influenza A virus-infected lung epithelial cells: the role of the nonstructural NS1 protein in the evasion of the host innate defense and its potential contribution to pandemic influenza. Proc Natl Acad Sci U S A 99:10736–10741 [View Article][PubMed]
    [Google Scholar]
  17. Goodman A. G., Smith J. A., Balachandran S., Perwitasari O., Proll S. C., Thomas M. J., Korth M. J., Barber G. N., Schiff L. A., Katze M. G. 2007; The cellular protein P58IPK regulates influenza virus mRNA translation and replication through a PKR-mediated mechanism. J Virol 81:2221–2230 [View Article][PubMed]
    [Google Scholar]
  18. Guo Z., Chen L. M., Zeng H., Gomez J. A., Plowden J., Fujita T., Katz J. M., Donis R. O., Sambhara S. 2007; NS1 protein of influenza A virus inhibits the function of intracytoplasmic pathogen sensor, RIG-I. Am J Respir Cell Mol Biol 36:263–269 [View Article][PubMed]
    [Google Scholar]
  19. Hale B. G., Steel J., Medina R. A., Manicassamy B., Ye J., Hickman D., Hai R., Schmolke M., Lowen A. C. et al. 2010; Inefficient control of host gene expression by the 2009 pandemic H1N1 influenza A virus NS1 protein. J Virol 84:6909–6922 [View Article][PubMed]
    [Google Scholar]
  20. Haye K., Burmakina S., Moran T., García-Sastre A., Fernandez-Sesma A. 2009; The NS1 protein of a human influenza virus inhibits type I interferon production and the induction of antiviral responses in primary human dendritic and respiratory epithelial cells. J Virol 83:6849–6862 [View Article][PubMed]
    [Google Scholar]
  21. Jackson D., Hossain M. J., Hickman D., Perez D. R., Lamb R. A. 2008; A new influenza virus virulence determinant: the NS1 protein four C-terminal residues modulate pathogenicity. Proc Natl Acad Sci U S A 105:4381–4386 [View Article][PubMed]
    [Google Scholar]
  22. Katze M. G., Krug R. M. 1990; Translational control in influenza virus-infected cells. Enzyme 44:265–277[PubMed]
    [Google Scholar]
  23. Katze M. G., Detjen B. M., Safer B., Krug R. M. 1986; Translational control by influenza virus: suppression of the kinase that phosphorylates the alpha subunit of initiation factor eIF-2 and selective translation of influenza viral mRNAs. Mol Cell Biol 6:1741–1750[PubMed]
    [Google Scholar]
  24. Katze M. G., Tomita J., Black T., Krug R. M., Safer B., Hovanessian A. 1988; Influenza virus regulates protein synthesis during infection by repressing autophosphorylation and activity of the cellular 68,000-M r protein kinase. J Virol 62:3710–3717[PubMed]
    [Google Scholar]
  25. Kochs G., García-Sastre A., Martínez-Sobrido L. 2007; Multiple anti-interferon actions of the influenza A virus NS1 protein. J Virol 81:7011–7021 [View Article][PubMed]
    [Google Scholar]
  26. Liu J., Lynch P. A., Chien C. Y., Montelione G. T., Krug R. M., Berman H. M. 1997; Crystal structure of the unique RNA-binding domain of the influenza virus NS1 protein. Nat Struct Biol 4:896–899 [View Article][PubMed]
    [Google Scholar]
  27. Liu H., Golebiewski L., Dow E. C., Krug R. M., Javier R. T., Rice A. P. 2010; The ESEV PDZ-binding motif of the avian influenza A virus NS1 protein protects infected cells from apoptosis by directly targeting Scribble. J Virol 84:11164–11174 [View Article][PubMed]
    [Google Scholar]
  28. Livak K. J., Schmittgen T. D. 2001; Analysis of relative gene expression data using real-time quantitative PCR and the . method. Methods 25:402–408 [View Article][PubMed]
    [Google Scholar]
  29. Lu Y., Wambach M., Katze M. G., Krug R. M. 1995; Binding of the influenza virus NS1 protein to double-stranded RNA inhibits the activation of the protein kinase that phosphorylates the eIF-2 translation initiation factor. Virology 214:222–228 [View Article][PubMed]
    [Google Scholar]
  30. Mibayashi M., Martínez-Sobrido L., Loo Y. M., Cárdenas W. B., Gale M. Jr, García-Sastre A. 2007; Inhibition of retinoic acid-inducible gene I-mediated induction of beta interferon by the NS1 protein of influenza A virus. J Virol 81:514–524 [View Article][PubMed]
    [Google Scholar]
  31. Nemeroff M. E., Barabino S. M., Li Y., Keller W., Krug R. M. 1998; Influenza virus NS1 protein interacts with the cellular 30 kDa subunit of CPSF and inhibits 3′ end formation of cellular pre-mRNAs. Mol Cell 1:991–1000 [View Article][PubMed]
    [Google Scholar]
  32. Noah D. L., Twu K. Y., Krug R. M. 2003; Cellular antiviral responses against influenza A virus are countered at the posttranscriptional level by the viral NS1A protein via its binding to a cellular protein required for the 3′ end processing of cellular pre-mRNAS. Virology 307:386–395 [View Article][PubMed]
    [Google Scholar]
  33. Nussbaum A. K., Rodriguez-Carreno M. P., Benning N., Botten J., Whitton J. L. 2005; Immunoproteasome-deficient mice mount largely normal CD8+ T cell responses to lymphocytic choriomeningitis virus infection and DNA vaccination. J Immunol 175:1153–1160[PubMed] [CrossRef]
    [Google Scholar]
  34. Obenauer J. C., Denson J., Mehta P. K., Su X., Mukatira S., Finkelstein D. B., Xu X., Wang J., Ma J. et al. 2006; Large-scale sequence analysis of avian influenza isolates. Science 311:1576–1580 [View Article][PubMed]
    [Google Scholar]
  35. Opitz B., Rejaibi A., Dauber B., Eckhard J., Vinzing M., Schmeck B., Hippenstiel S., Suttorp N., Wolff T. 2007; IFNβ induction by influenza A virus is mediated by RIG-I which is regulated by the viral NS1 protein. Cell Microbiol 9:930–938 [View Article][PubMed]
    [Google Scholar]
  36. Park M. S., Shaw M. L., Muñoz-Jordan J., Cros J. F., Nakaya T., Bouvier N., Palese P., García-Sastre A., Basler C. F. 2003; Newcastle disease virus (NDV)-based assay demonstrates interferon-antagonist activity for the NDV V protein and the Nipah virus V, W, and C proteins. J Virol 77:1501–1511 [View Article][PubMed]
    [Google Scholar]
  37. Pichlmair A., Schulz O., Tan C. P., Näslund T. I., Liljeström P., Weber F., Reis e Sousa C. 2006; RIG-I-mediated antiviral responses to single-stranded RNA bearing 5′-phosphates. Science 314:997–1001 [View Article][PubMed]
    [Google Scholar]
  38. Quinlivan M., Zamarin D., García-Sastre A., Cullinane A., Chambers T., Palese P. 2005; Attenuation of equine influenza viruses through truncations of the NS1 protein. J Virol 79:8431–8439 [View Article][PubMed]
    [Google Scholar]
  39. Seifert U., Bialy L. P., Ebstein F., Bech-Otschir D., Voigt A., Schröter F., Prozorovski T., Lange N., Steffen J. et al. 2010; Immunoproteasomes preserve protein homeostasis upon interferon-induced oxidative stress. Cell 142:613–624 [View Article][PubMed]
    [Google Scholar]
  40. Solórzano A., Webby R. J., Lager K. M., Janke B. H., García-Sastre A., Richt J. A. 2005; Mutations in the NS1 protein of swine influenza virus impair anti-interferon activity and confer attenuation in pigs. J Virol 79:7535–7543 [View Article][PubMed]
    [Google Scholar]
  41. Steel J., Burmakina S. V., Thomas C., Spackman E., García-Sastre A., Swayne D. E., Palese P. 2008; A combination in-ovo vaccine for avian influenza virus and Newcastle disease virus. Vaccine 26:522–531 [View Article][PubMed]
    [Google Scholar]
  42. Thanos D., Maniatis T. 1995; Virus induction of human IFN β gene expression requires the assembly of an enhanceosome. Cell 83:1091–1100 [View Article][PubMed]
    [Google Scholar]
  43. Thompson W. W., Shay D. K., Weintraub E., Brammer L., Cox N., Anderson L. J., Fukuda K. 2003; Mortality associated with influenza and respiratory syncytial virus in the United States. JAMA 289:179–186 [View Article][PubMed]
    [Google Scholar]
  44. Thompson W. W., Shay D. K., Weintraub E., Brammer L., Bridges C. B., Cox N. J., Fukuda K. 2004; Influenza-associated hospitalizations in the United States. JAMA 292:1333–1340 [View Article][PubMed]
    [Google Scholar]
  45. Wang X., Li M., Zheng H., Muster T., Palese P., Beg A. A., García-Sastre A. 2000; Influenza A virus NS1 protein prevents activation of NF-κB and induction of alpha/beta interferon. J Virol 74:11566–11573 [View Article][PubMed]
    [Google Scholar]
  46. Zielecki F., Semmler I., Kalthoff D., Voss D., Mauel S., Gruber A. D., Beer M., Wolff T. 2010; Virulence determinants of avian H5N1 influenza A virus in mammalian and avian hosts: role of the C-terminal ESEV motif in the viral NS1 protein. J Virol 84:10708–10718 [View Article][PubMed]
    [Google Scholar]
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