1887

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Volume 90, Issue 2

Effects of multiple amino acids of the parainfluenza virus 5 fusion protein on its haemagglutinin–neuraminidase-independent fusion activity Free

Abstract

The fusion (F) protein of parainfluenza virus 5 (PIV-5) strain W3A is able to induce cell fusion when it is expressed alone in baby hamster kidney cells, whilst the F protein of PIV-5 strain WR induces cell fusion only when co-expressed with the haemagglutinin–neuraminidase (HN) protein. It has been shown previously that when Leu-22 of the WR F protein is replaced with the W3A F counterpart (Pro-22), the resulting mutant L22P exhibits HN-independent fusion activity. Furthermore, previous chimeric analysis between L22P and the F protein of PIV-5 strain T1 has suggested that Glu-132 also contributes to the HN-independent fusion activity of L22P. It was shown here that substitution of Glu-132 of L22P with various amino acids including the T1 F protein counterpart (Lys-132) resulted in a reduction in fusion activity, whereas substitution with Asp was the exception in being tolerated. Interestingly, reduced fusion activity of an L22P mutant that harboured the E132K substitution could be restored by an additional D416K substitution but not by a D416E mutation, suggesting that the presence of the same charge at positions 132 and 416 is important for the HN-independent fusion activity. In contrast, substitution of Leu-22 of the WR F protein with various amino acids except those with aliphatic side chains resulted in acquisition of fusion activity, suggesting that the HN dependence of the WR F protein in the induction of cell fusion is attributable to the hydrophobicity of Leu-22. These results indicate that at least three amino acids are involved in the HN-independent fusion activity of the PIV-5 F protein.

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2009-02-01
2024-04-27
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References

  1. Deng R., Mirza A. M., Mahon P. J., Iorio R. M. 1997; Functional chimeric HN glycoproteins derived from Newcastle disease virus and human parainfluenza virus-3. Arch Virol Suppl 13:115–130
     [Google Scholar]
  2. Dutch R. E., Hagglund R. N., Nagel M. A., Paterson R. G., Lamb R. A. 2001; Paramyxovirus fusion (F) protein: a conformational change on cleavage activation. Virology 281:138–150 [CrossRef]
     [Google Scholar]
  3. Gething M.-J., White J. M., Waterfield M. D. 1978; Purification of the fusion protein of Sendai virus: analysis of the NH2-terminal sequence generated during precursor activation. Proc Natl Acad Sci U S A 75:2737–2740 [CrossRef]
     [Google Scholar]
  4. Graham F. L., van der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467 [CrossRef]
     [Google Scholar]
  5. Horvath C. M., Paterson R. G., Shaughnessy M. A., Wood R., Lamb R. A. 1992; Biological activity of paramyxovirus fusion proteins: factors influencing formation of syncytia. J Virol 66:4564–4569
     [Google Scholar]
  6. Hsu M.-C., Scheid A., Choppin P. W. 1981; Activation of the Sendai virus fusion protein (F) involved a conformational change with exposure of a new hydrophobic region. J Biol Chem 256:3557–3565
     [Google Scholar]
  7. Hu X., Ray R., Compans R. W. 1992; Functional interactions between the fusion protein and hemagglutinin-neuraminidase of human parainfluenza viruses. J Virol 66:1528–1534
     [Google Scholar]
  8. Ito M., Nishio M., Kawano M., Kusagawa S., Komada H., Ito Y., Tsurudome M. 1997; Role of a single amino acid at the amino terminus of the simian virus 5 F2 subunit in syncytium formation. J Virol 71:9855–9858
     [Google Scholar]
  9. Ito M., Nishio M., Komada H., Ito Y., Tsurudome M. 2000; An amino acid in the heptad repeat 1 domain is important for the haemagglutinin–neuraminidase-independent fusing activity of simian virus 5 fusion protein. J Gen Virol 81:719–727
     [Google Scholar]
  10. Karron R. A., Collins P. L. 2007; Parainfluenza viruses. In Fields Virology , 5th edn. pp 1497–1526Edited by Knipe D. M., Howley P. M., Griffin D. E., Martin M. A., Lamb R. A., Roizman B., Straus S. E. Philadelphia, PA: Lippincott Williams & Wilkins;
     [Google Scholar]
  11. Kohama T., Garten W., Klenk H.-D. 1981; Changes in conformation and charge paralleling proteolytic activation of Newcastle disease virus glycoproteins. Virology 111:364–376 [CrossRef]
     [Google Scholar]
  12. Lamb R. A., Parks G. D. 2007; Paramyxoviridae : the viruses and their replication. In Fields Virology , 5th edn. pp 1449–1496Edited by Knipe D. M., Howley P. M., Griffin D. E., Martin M. A., Lamb R. A., Roizman B., Straus S. E. Philadelphia, PA: Lippincott Williams & Wilkins;
     [Google Scholar]
  13. Lee J. K., Prussia A., Paal T., White L. K., Snyder J. P., Plemper R. K. 2008; Functional interaction between paramyxovirus fusion and attachment proteins. J Biol Chem 283:16561–16572 [CrossRef]
     [Google Scholar]
  14. Ludwig K., Schade B., Böttcher C., Korte T., Ohlwein N., Baljinnyam B., Veit M., Herrmann A. 2008; Electron cryomicroscopy reveals different F1+F2 protein states in intact parainfluenza virions. J Virol 82:3775–3781 [CrossRef]
     [Google Scholar]
  15. McGinnes L. W., Morrison T. G. 2006; Inhibition of receptor binding stabilizes Newcastle disease virus HN and F protein-containing complexes. J Virol 80:2894–2903 [CrossRef]
     [Google Scholar]
  16. Melanson V. R., Iorio R. M. 2006; Addition of N -glycans in the stalk of the Newcastle disease virus HN protein blocks its interaction with the F protein and prevents fusion. J Virol 80:623–633 [CrossRef]
     [Google Scholar]
  17. Novick S. L., Hoekstra D. 1988; Membrane penetration of Sendai virus glycoproteins during the early stages of fusion with liposomes as determined by hydrophobic photoaffinity labelling. Proc Natl Acad Sci U S A 85:7433–7437 [CrossRef]
     [Google Scholar]
  18. Paterson R. G., Hiebert S. W., Lamb R. A. 1985; Expression at the cell surface of biologically active fusion and hemagglutinin/neuraminidase proteins of the paramyxovirus simian virus 5 from cloned cDNA. Proc Natl Acad Sci U S A 82:7520–7524 [CrossRef]
     [Google Scholar]
  19. Paterson R. G., Shaughnessy M. A., Lamb R. A. 1989; Analysis of the relationship between cleavability of a paramyxovirus fusion protein and length of the connecting peptide. J Virol 63:1293–1301
     [Google Scholar]
  20. Paterson R. G., Russell C. J., Lamb R. A. 2000; Fusion protein of the paramyxovirus SV5: destabilizing and stabilizing mutants of fusion activation. Virology 270:17–30 [CrossRef]
     [Google Scholar]
  21. Russell C. J., Jardetzky T. S., Lamb R. A. 2004; Conserved glycine residues in the fusion peptide of the paramyxovirus fusion protein regulate activation of the native state. J Virol 78:13727–13742 [CrossRef]
     [Google Scholar]
  22. Sakata H., Hishiyama M., Sugiura A. 1984; Enzyme-linked immunosorbent assay compared with neutralization tests for evaluation of live mumps vaccines. J Clin Microbiol 19:21–25
     [Google Scholar]
  23. Sergel T. A., McGinnes L. W., Morrison T. G. 2000; A single amino acid change in the Newcastle disease virus fusion protein alters the requirement for HN protein in fusion. J Virol 74:5101–5107 [CrossRef]
     [Google Scholar]
  24. Seth S., Vincent A., Compans R. W. 2003; Mutations in the cytoplasmic domain of a paramyxovirus fusion glycoprotein rescue syncytium formation and eliminate the hemagglutinin–neuraminidase protein requirement for membrane fusion. J Virol 77:167–178 [CrossRef]
     [Google Scholar]
  25. Takimoto T., Taylor G. L., Connaris H. C., Crennell S. J., Portner A. 2002; Role of the hemagglutinin–neuraminidase protein in the mechanism of paramyxovirus–cell membrane fusion. J Virol 76:13028–13033 [CrossRef]
     [Google Scholar]
  26. Tanabayashi K., Compans R. W. 1996; Functional interaction of paramyxovirus glycoproteins: identification of a domain in Sendai virus HN which promotes cell fusion. J Virol 70:6112–6118
     [Google Scholar]
  27. Tong S., Li M., Vincent A., Compans R. W., Fritsch E., Beier R., Klenk C., Ohuchi M., Klenk H.-D. 2002; Regulation of fusion activity by the cytoplasmic domain of a paramyxovirus F protein. Virology 301:322–333 [CrossRef]
     [Google Scholar]
  28. Tsurudome M., Kawano M., Yuasa T., Tabata N., Nishio M., Komada H., Ito Y. 1995; Identification of regions on the hemagglutinin–neuraminidase protein of human parainfluenza virus type 2 important for promoting cell fusion. Virology 213:190–203 [CrossRef]
     [Google Scholar]
  29. Tsurudome M., Ito M., Nishio M., Kawano M., Okamoto K., Kusagawa S., Komada H., Ito Y. 1998; Identification of regions on the fusion protein of human parainfluenza virus type 2 which are required for haemagglutinin–neuraminidase proteins to promote cell fusion. J Gen Virol 79:279–289
     [Google Scholar]
  30. Tsurudome M., Ito M., Nishio M., Kawano M., Komada H., Ito Y. 2001; Hemagglutinin–neuraminidase-independent fusion activity of simian virus 5 fusion (F) protein: difference in conformation between fusogenic and nonfusogenic F proteins on the cell surface. J Virol 75:8999–9009 [CrossRef]
     [Google Scholar]
  31. Tsurudome M., Ito M., Nishio M., Kawano M., Komada H., Ito Y. 2006; A mutant fusion (F) protein of simian virus 5 induces hemagglutinin–neuraminidase-independent syncytium formation despite the internalization of the F protein. Virology 347:11–27 [CrossRef]
     [Google Scholar]
  32. Umino Y., Kohama T., Sato T. A., Sugiura A., Klenk H.-D., Rott R. 1990; Monoclonal antibodies to three structural proteins of Newcastle disease virus: biological characterization with particular reference to the conformational change of envelope glycoproteins associated with proteolytic cleavage. J Gen Virol 71:1189–1197 [CrossRef]
     [Google Scholar]
  33. Yin H. S., Paterson R. G., Wen X., Lamb R. A., Jardetzky T. S. 2005; Structure of the uncleaved ectodomain of the paramyxovirus (hPIV3) fusion protein. Proc Natl Acad Sci U S A 102:9288–9293 [CrossRef]
     [Google Scholar]
  34. Yin H. S., Wen X., Paterson R. G., Lamb R. A., Jardetzky T. S. 2006; Structure of the parainfluenza virus 5 F protein in its metastable, prefusion conformation. Nature 439:38–44 [CrossRef]
     [Google Scholar]
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Effects of multiple amino acids of the parainfluenza virus 5 fusion protein on its haemagglutinin–neuraminidase-independent fusion activity
J. Gen. Virol. 90, 405 (2009); https://doi.org/10.1099/vir.0.006437-0
/content/journal/jgv/10.1099/vir.0.006437-0
/content/journal/jgv/10.1099/vir.0.006437-0
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