1887

Abstract

In this study we compared the NS1 protein of Influenza B/Lee/40 and several non-cultured Influenza B virus clinical strains detected in Singapore. In B/Lee/40 virus-infected cells and in cells expressing the recombinant B/Lee/40 NS1 protein a full-length 35 kDa NS1 protein and a 23 kDa NS1 protein species (p23) were detected. Mutational analysis of the NS1 gene indicated that p23 was generated by a novel cleavage event within the linker domain between an aspartic acid and proline at amino acid residues at positions 92 and 93 respectively (DP92–93), and that p23 contained the first 92 amino acids of the NS1 protein. Sequence analysis of the Singapore strains indicated the presence of either DP92–93 or NP92–93 in the NS1 protein, but protein expression analysis showed that p23 was only detected in NS1 proteins with DP92–93.. An additional adjacent proline residue at position 94 (P94) was present in some strains and correlated with increased p23 levels, suggesting that P94 has a synergistic effect on the cleavage of the NS1 protein. The first 145 amino acids of the NS1 protein are required for inhibition of ISG15-mediated ubiquitination, and our analysis showed that Influenza B viruses circulating in Singapore with DP92–93 expressed truncated NS1 proteins and may differ in their capacity to inhibit ISG15 activity. Thus, DP92–93 in the NS1 protein may confer a disadvantage to Influenza B viruses circulating in the human population and interestingly the low frequency of DP92–93detection in the NS1 protein since 2004 is consistent with this suggestion.

Keyword(s): B/Lee/40 , Cleavage , Influenza B , NS1 and Proteolysis
Loading

Article metrics loading...

/content/journal/mgen/10.1099/mgen.0.000082
2016-08-25
2020-12-01
Loading full text...

Full text loading...

/deliver/fulltext/mgen/2/8/mgen000082.html?itemId=/content/journal/mgen/10.1099/mgen.0.000082&mimeType=html&fmt=ahah

References

  1. Alnemri E. S., Livingston D. J., Nicholson D. W., Salvesen G., Thornberry N. A., Wong W. W., Yuan J.. 1996; Human ICE/CED-3 protease nomenclature. Cell87:171 [CrossRef][PubMed]
    [Google Scholar]
  2. Baine W. B., Luby J. P., Martin S. M.. 1980; Severe illness with influenza B. Am J Med68:181–189 [CrossRef][PubMed]
    [Google Scholar]
  3. Briedis D. J., Lamb R. A.. 1982; Influenza B virus genome: sequences and structural organization of RNA segment 8 and the mRNAs coding for the NS1 and NS2 proteins. J Virol42:186–193[PubMed]
    [Google Scholar]
  4. Dauber B., Heins G., Wolff T.. 2004; The influenza B virus nonstructural NS1 protein is essential for efficient viral growth and antagonizes beta interferon induction. J Virol78:1865–1872 [CrossRef][PubMed]
    [Google Scholar]
  5. Dauber B., Schneider J., Wolff T.. 2006; Double-stranded RNA binding of influenza B virus nonstructural NS1 protein inhibits protein kinase R but is not essential to antagonize production of alpha/beta interferon. J Virol80:11667–11677 [CrossRef][PubMed]
    [Google Scholar]
  6. Derlet R. W.. 2010; Influenza. eMedicine Infectious Diseases
    [Google Scholar]
  7. Donelan N. R., Dauber B., Wang X., Basler C. F., Wolff T., García-Sastre A.. 2004; The N- and C-terminal domains of the NS1 protein of influenza B virus can independently inhibit IRF-3 and beta interferon promoter activation. J Virol78:11574–11582 [CrossRef][PubMed]
    [Google Scholar]
  8. Guan R., Ma L.-C., Leonard P. G., Amer B. R., Sridharan H., Zhao C., Krug R. M., Montelione G. T.. 2011; Structural basis for the sequence-specific recognition of human ISG15 by the NS1 protein of influenza B virus. Proc Natl Acad Sci U S A108:13468–13473 [CrossRef]
    [Google Scholar]
  9. Hosur M. V., Schmidt T., Tucker R. C., Johnson J. E., Gallagher T. M., Selling B. H., Rueckert R. R.. 1987; Structure of an insect virus at 3.0 Å resolution. Proteins2:167–176 [CrossRef][PubMed]
    [Google Scholar]
  10. Jumat M. R., Sugrue R. J., Tan B. H.. 2014; Genetic characterisation of influenza B viruses detected in Singapore, 2004 to 2009. BMC Res Notes7: [CrossRef][PubMed]
    [Google Scholar]
  11. Kim Y. H., Kim H. S., Cho S. H., Seo S. H.. 2009; Influenza B virus causes milder pathogenesis and weaker inflammatory responses in ferrets than influenza A virus. Viral Immunol22:423–430 [CrossRef][PubMed]
    [Google Scholar]
  12. Li L., Wang D., Jiang Y., Sun J., Zhang S., Chen Y., Wang X.. 2011; Crystal structure of human ISG15 protein in complex with influenza B virus NS1B. J Biol Chem286:30258–30262 [CrossRef][PubMed]
    [Google Scholar]
  13. Lüthi A. U., Martin S. J.. 2007; The CASBAH: a searchable database of caspase substrates. Cell Death Differ14:641–650 [CrossRef][PubMed]
    [Google Scholar]
  14. Michael K. F., John C. P., Kevin E. B., Robert J., Pamela K. H.. 1980; Epidemic influenza myopathy in Cincinnati in 1977. J Pediatr96:545–551[PubMed]
    [Google Scholar]
  15. Muramoto Y., Noda T., Kawakami E., Akkina R., Kawaoka Y.. 2013; Identification of novel influenza A virus proteins translated from PA mRNA. J Virol87:2455–2462 [CrossRef][PubMed]
    [Google Scholar]
  16. Patch J. R., Crameri G., Wang L. F., Eaton B. T., Broder C. C.. 2007; Quantitative analysis of Nipah virus proteins released as virus-like particles reveals central role for the matrix protein. Virol J4:1 [CrossRef][PubMed]
    [Google Scholar]
  17. Randall R. E., Goodbourn S.. 2008; Interferons and viruses: an interplay between induction, signalling, antiviral responses and virus countermeasures. J Gen Virol89:1–47 [CrossRef][PubMed]
    [Google Scholar]
  18. Richard A., Tulasne D.. 2012; Caspase cleavage of viral proteins, another way for viruses to make the best of apoptosis. Cell Death Dis3:e277 [CrossRef][PubMed]
    [Google Scholar]
  19. Schneider J., Dauber B., Melén K., Julkunen I., Wolff T.. 2009; Analysis of influenza B Virus NS1 protein trafficking reveals a novel interaction with nuclear speckle domains. J Virol83:701–711 [CrossRef][PubMed]
    [Google Scholar]
  20. Shaw M., Peter P.. 2013; Orthomyxoviridae. Fields Virology, 6th.1151
    [Google Scholar]
  21. Speir J. A., Taylor D. J., Natarajan P., Pringle F. M., Ball L. A., Johnson J. E.. 2010; Evolution in action: N and C termini of subunits in related T = 4 viruses exchange roles as molecular switches. Structure18:700–709 [CrossRef][PubMed]
    [Google Scholar]
  22. Sridharan H., Zhao C., Krug R. M.. 2010; Species specificity of the NS1 protein of influenza B virus: NS1 binds only human and non-human primate ubiquitin-like ISG15 proteins. J Biol Chem285: [CrossRef][PubMed]
    [Google Scholar]
  23. Talon J., Salvatore M., O'Neill R. E., Nakaya Y., Zheng H., Muster T., García-Sastre A., Palese P.. 2000; Influenza A and B viruses expressing altered NS1 proteins: A vaccine approach. Proc Natl Acad Sci U S A97:4309–4314 [CrossRef][PubMed]
    [Google Scholar]
  24. Tauber S., Ligertwood Y., Quigg-Nicol M., Dutia B. M., Elliott R. M.. 2012; Behaviour of influenza A viruses differentially expressing segment 2 gene products in vitro and in vivo. J Gen Virol93:840–849 [CrossRef][PubMed]
    [Google Scholar]
  25. Tobita K., Tanaka T., Odagiri T., Tashiro M., Feng S. Y.. 1990; Nucleotide sequence and some biological properties of the NS gene of a newly isolated influenza B virus mutant which has a long carboxyl terminal deletion in the NS1 protein. Virology174:314–319 [CrossRef][PubMed]
    [Google Scholar]
  26. Wang W., Krug R. M.. 1996; The RNA-binding and effector domains of the viral NS1 protein are conserved to different extents among influenza A and B viruses. Virology223:41–50 [CrossRef][PubMed]
    [Google Scholar]
  27. Wery J. P., Reddy V. S., Hosur M. V., Johnson J. E.. 1994; The refined three-dimensional structure of an insect virus at 2.8 Å resolution. J Mol Biol235:565–586 [CrossRef][PubMed]
    [Google Scholar]
  28. Wise H. M., Foeglein A., Sun J., Dalton R. M., Patel S., Howard W., Anderson E. C., Barclay W. S., Digard P.. 2009; A complicated message: Identification of a novel PB1-related protein translated from influenza A virus segment 2 mRNA. J Virol83:8021–8031 [CrossRef][PubMed]
    [Google Scholar]
  29. World Health Organization 2014; Influenza (Seasonal) Fact Sheet Number 211. Geneva: WHO;
  30. Wu C.-T., Hsia S.-H., Huang J.-L.. 2009; Influenza B-associated rhabdomyolysis in Taiwanese children. Acta Paediatrica99:1701–1704 [CrossRef]
    [Google Scholar]
  31. Yuan W., Krug R. M.. 2001; Influenza B virus NS1 protein inhibits conjugation of the interferon (IFN)-induced ubiquitin-like ISG15 protein. EMBO J20:362–371 [CrossRef][PubMed]
    [Google Scholar]
  32. Zhao C., Collins M. N., Hsiang T. Y., Krug R. M.. 2013; Interferon-induced ISG15 pathway: an ongoing virus-host battle. Trends Microbiol21:181–186 [CrossRef][PubMed]
    [Google Scholar]
  33. Zhirnov O. P., Konakova T. E., Garten W., Klenk H.. 1999; Caspase-dependent N-terminal cleavage of influenza virus nucleocapsid protein in infected cells. J Virol73:10158–10163[PubMed]
    [Google Scholar]
  34. Zhirnov O. P., Konakova T. E., Wolff T., Klenk H. D.. 2002; NS1 protein of influenza A virus down-regulates apoptosis. J Virol76:1617–1625 [CrossRef][PubMed]
    [Google Scholar]
  35. Jumat, M. R., Sugrue, R. J. & Tan, B. H., GenBank, Influenza B virus nuclear export protein (NEP) gene, partial cds; and nonstructural protein 1 (NS1) gene, complete cds. PopSet: 482577243 KC844161-KC844196 2014
  36. Jumat, M. R., Sugrue, R. J. & Tan, B. H., GenBankGenBank, Influenza B virus (B/Singapore/DSO_090136/2004) segment 8 nuclear export protein (NEP) and nonstructural protein 1 (NS1) genes, complete cds. GenBankGenBank: KC844195.1 2016
  37. Jumat, M. R., Sugrue, R. J. & Tan, B. H., GenBankGenBank, Influenza B virus (B/Singapore/DSO_040117Variant/2006) segment 8 NS1 gene, complete sequence; nonstructural protein 1 (NEP) gene, complete cds; and nuclear export protein (NS2) gene, partial cds, GenBankGenBank: KU500812.1 2016
  38. Jumat, M. R., Sugrue, R. J. & Tan, B. H., GenBankGenBank, Influenza B virus (B/Singapore/DSO_010147/2007) segment 8 nuclear export protein (NEP) and nonstructural protein 1 (NS1) genes, complete cds, GenBankGenBank: KC844165.1 2016
  39. Jumat, M. R., Sugrue, R. J. & Tan, B. H., GenBankGenBank, Influenza B virus (B/Singapore/DSO_020132/2007) segment 8 nuclear export protein (NEP) and nonstructural protein 1 (NS1) genes, complete cds, GenBankGenBank: KC844167.1 2016
  40. Jumat, M. R., Sugrue, R. J. & Tan, B. H., GenBankGenBank, Influenza B virus (B/Singapore/DSO_0070/2009) segment 8 nuclear export protein (NEP) gene, partial cds; and nonstructural protein 1 (NS1) gene, complete cds, GenBankGenBank: KC844163.1 2016
http://instance.metastore.ingenta.com/content/journal/mgen/10.1099/mgen.0.000082
Loading
/content/journal/mgen/10.1099/mgen.0.000082
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