1887

Abstract

The cellular distribution of the small hydrophobic (SH) protein in respiratory syncytial virus (RSV)-infected cells was examined. Although the SH protein was distributed throughout the cytoplasm, it appeared to accumulate in the Golgi complex within membrane structures that were enriched in the raft lipid, GM1. The ability of the SH protein to interact with lipid-raft membranes was further confirmed by examining its detergent-solubility properties in Triton X-100 at 4 °C. This analysis showed that a large proportion of the SH protein exhibited detergent-solubility characteristics that were consistent with an association with lipid-raft membranes. Analysis of virus-infected cells by immuno-transmission electron microscopy revealed SH protein clusters on the cell surface, but only very low levels of the protein appeared to be associated with mature virus filaments and inclusion bodies. These data suggest that during virus infection, the compartments in the secretory pathway, such as the endoplasmic reticulum (ER) and Golgi complex, are major sites of accumulation of the SH protein. Furthermore, although a significant amount of this protein interacts with lipid-raft membranes within the Golgi complex, its presence within mature virus filaments is minimal.

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2004-05-01
2019-11-22
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References

  1. Anderson, K., King, A. M., Lerch, R. A. & Wertz, G. W. ( 1992; ). Polylactosaminoglycan modification of the respiratory syncytial virus small hydrophobic (SH) protein: a conserved feature among human and bovine respiratory syncytial viruses. Virology 191, 417–430.[CrossRef]
    [Google Scholar]
  2. Brown, D. A. & London, E. ( 1998; ). Functions of lipid rafts in biological membranes. Annu Rev Cell Dev Biol 14, 111–136.[CrossRef]
    [Google Scholar]
  3. Brown, G., Aitken, J., Rixon, H. W. McL. & Sugrue, R. J. ( 2002a; ). Caveolin-1 is incorporated into mature respiratory syncytial virus particles during virus assembly on the surface of virus-infected cells. J Gen Virol 83, 611–621.
    [Google Scholar]
  4. Brown, G., Rixon, H. W. McL. & Sugrue, R. J. ( 2002b; ). Respiratory syncytial virus assembly occurs in GM1-rich regions of the host-cell membrane and alters the cellular distribution of tyrosine phosphorylated caveolin-1. J Gen Virol 83, 1841–1850.
    [Google Scholar]
  5. Bukreyev, A., Whitehead, S. S., Murphy, B. R. & Collins, P. L. ( 1997; ). Recombinant respiratory syncytial virus from which the entire SH gene has been deleted grows efficiently in cell culture and exhibits site-specific attenuation in the respiratory tract of the mouse. J Virol 71, 8973–8982.
    [Google Scholar]
  6. Chen, M. D., Vazquez, M., Buonocore, L. & Kahn, J. S. ( 2000; ). Conservation of the respiratory syncytial virus SH gene. J Infect Dis 182, 1228–1233.[CrossRef]
    [Google Scholar]
  7. Ciampor, F., Bayley, P. M., Nermut, M. V., Hirst, E. M. A., Sugrue, R. J. & Hay, A. J. ( 1992; ). Evidence that the amantadine-induced M2-mediated conversion of influenza A virus hemagglutinin to the low pH conformation occurs in an acidic trans Golgi compartment. Virology 188, 14–24.[CrossRef]
    [Google Scholar]
  8. Collins, P. L. & Wertz, G. W. ( 1985; ). The 1A protein gene of human respiratory syncytial virus: nucleotide sequence of the mRNA and a related polycistronic transcript. Virology 141, 283–291.[CrossRef]
    [Google Scholar]
  9. Collins, P. L. & Mottet, G. ( 1991; ). Post-translational processing and oligomerization of the fusion glycoprotein of human respiratory syncytial virus. J Gen Virol 72, 3095–3101.[CrossRef]
    [Google Scholar]
  10. Collins, P. L. & Mottet, G. ( 1993; ). Membrane orientation and oligomerization of the small hydrophobic protein of human respiratory syncytial virus. J Gen Virol 74, 1445–1450.[CrossRef]
    [Google Scholar]
  11. Collins, P. L., Olmsted, R. A. & Johnson, P. R. ( 1990; ). The small hydrophobic protein of human respiratory syncytial virus: comparison between antigenic subgroups A and B. J Gen Virol 71, 1571–1576.[CrossRef]
    [Google Scholar]
  12. Feldman, S. A., Crim, R. L., Audet, S. A. & Beeler, J. A. ( 2001; ). Human respiratory syncytial virus surface glycoproteins F, G and SH form an oligomeric complex. Arch Virol 146, 2369–2383.[CrossRef]
    [Google Scholar]
  13. Garcia, J., Garcia-Barreno, B., Vivo, A. & Melero, J. A. ( 1993; ). Cytoplasmic inclusions of respiratory syncytial virus-infected cells: formation of inclusion bodies in transfected cells that coexpress the nucleoprotein, the phosphoprotein, and the 22K protein. Virology 195, 243–247.[CrossRef]
    [Google Scholar]
  14. Garcia-Barreno, B., Delgado, T. & Melero, J. A. ( 1996; ). Identification of protein regions involved in the interaction of human respiratory syncytial virus phosphoprotein and nucleoprotein: significance for nucleocapsid assembly and formation of cytoplasmic inclusions. J Virol 70, 801–808.
    [Google Scholar]
  15. Ghildyal, R., Mills, J., Murray, M., Vardaxis, N. & Meanger, J. ( 2002; ). Respiratory syncytial virus matrix protein associates with nucleocapsids in infected cells. J Gen Virol 83, 753–757.
    [Google Scholar]
  16. Heminway, B. R., Yu, Y., Tanaka, Y., Perrine, K. G., Gustafson, E., Bernstein, J. M. & Galinski, M. S. ( 1994; ). Analysis of respiratory syncytial virus F, G, and SH proteins in cell fusion. Virology 200, 801–805.[CrossRef]
    [Google Scholar]
  17. Henderson, G., Murray, J. & Yeo, R. P. ( 2002; ). Sorting of the respiratory syncytial virus matrix protein into detergent-resistant structures is dependent on cell-surface expression of the glycoproteins. Virology 300, 244–254.[CrossRef]
    [Google Scholar]
  18. Henkel, J. R. & Weisz, O. A. ( 1998; ). Influenza virus M2 protein slows traffic along the secretory pathway. pH perturbation of acidified compartments affects early Golgi transport steps. J Biol Chem 273, 6518–6524.[CrossRef]
    [Google Scholar]
  19. Henkel, J. R., Apodaca, G., Altschuler, Y., Hardy, S. & Weisz, O. A. ( 1998; ). Selective perturbation of apical membrane traffic by expression of influenza M2, an acid-activated ion channel, in polarized Madin–Darby canine kidney cells. Mol Biol Cell 9, 2477–2490.[CrossRef]
    [Google Scholar]
  20. Henkel, J. R., Gibson, G. A., Poland, P. A., Ellis, M. A., Hughey, R. P. & Weisz, O. A. ( 2000; ). Influenza M2 proton channel activity selectively inhibits trans-Golgi network release of apical membrane and secreted proteins in polarized Madin–Darby canine cells. J Cell Biol 148, 495–504.[CrossRef]
    [Google Scholar]
  21. Ikonen, E. ( 2001; ). Roles of lipid rafts in membrane transport. Curr Opin Cell Biol 13, 470–477.[CrossRef]
    [Google Scholar]
  22. Jeffree, C. E., Rixon, H. W. McL., Brown, G., Aitken, J. & Sugrue, R. J. ( 2003; ). Distribution of the attachment (G) glycoprotein and GM1 within the envelope of mature respiratory syncytial virus filaments revealed using field emission scanning electron microscopy. Virology 306, 254–267.[CrossRef]
    [Google Scholar]
  23. Le, P. U. & Nabi, I. R. ( 2003; ). Distinct caveolae-mediated endocytic pathways target the Golgi apparatus and the endoplasmic reticulum. J Cell Sci 116, 1059–1071.[CrossRef]
    [Google Scholar]
  24. Lencer, W. I., Delp, C., Neutra, M. R. & Madara, J. L. ( 1992; ). Mechanism of cholera toxin action on a polarized human intestinal epithelial cell line: role of vesicular traffic. J Cell Biol 117, 1197–1209.[CrossRef]
    [Google Scholar]
  25. Lencer, W. I., Constable, C., Moe, S., Jobling, M. G., Webb, H. M., Ruston, S., Madara, J. L., Hirst, T. R. & Holmes, R. K. ( 1995; ). Targeting of cholera toxin and Escherichia coli heat labile toxin in polarized epithelia: role of COOH-terminal KDEL. J Cell Biol 131, 951–962.[CrossRef]
    [Google Scholar]
  26. Lencer, W. I., Hirst, T. R. & Holmes, R. K. ( 1999; ). Membrane traffic and the cellular uptake of cholera toxin. Biochim Biophys Acta 1450, 177–190.[CrossRef]
    [Google Scholar]
  27. McCurdy, L. H. & Graham, B. S. ( 2003; ). Role of plasma membrane lipid microdomains in respiratory syncytial virus filament formation. J Virol 77, 1747–1756.[CrossRef]
    [Google Scholar]
  28. Mould, J. A., Paterson, R. G., Takeda, M., Ohigashi, Y., Venkataraman, P., Lamb, R. A. & Pinto, L. H. ( 2003; ). Influenza B virus BM2 protein has ion channel activity that conducts protons across membranes. Dev Cell 5, 175–184.[CrossRef]
    [Google Scholar]
  29. Olmsted, R. A. & Collins, P. L. ( 1989; ). The 1A protein of respiratory syncytial virus is an integral membrane protein present as multiple, structurally distinct species. J Virol 63, 2019–2029.
    [Google Scholar]
  30. Parry, J. E., Shirodaria, P. V. & Pringle, C. R. ( 1979; ). Pneumoviruses: the cell surface of lytically and persistently infected cells. J Gen Virol 44, 479–491.[CrossRef]
    [Google Scholar]
  31. Perez, M., Garcia-Barreno, B., Melero, J. A., Carrasco, L. & Guinea, R. ( 1997; ). Membrane permeability changes induced in Escherichia coli by the SH protein of human respiratory syncytial virus. Virology 235, 342–351.[CrossRef]
    [Google Scholar]
  32. Rixon, H. W., Brown, C., Brown, G. & Sugrue, R. J. ( 2002; ). Multiple glycosylated forms of the respiratory syncytial virus fusion protein are expressed in virus-infected cells. J Gen Virol 83, 61–66.
    [Google Scholar]
  33. Roberts, S. R., Compans, R. W. & Wertz, G. W. ( 1995; ). Respiratory syncytial virus matures at the apical surfaces of polarized epithelial cells. J Virol 69, 2667–2673.
    [Google Scholar]
  34. Sakaguchi, T., Leser, G. P. & Lamb, R. A. ( 1996; ). The ion channel activity of the influenza virus M2 protein affects transport through the Golgi apparatus. J Cell Biol 133, 733–747.[CrossRef]
    [Google Scholar]
  35. Sugrue, R. J., Bahadur, G., Zambon, M. C., Hall-Smith, M., Douglas, A. R. & Hay, A. J. ( 1990; ). Specific structural alteration of the influenza haemagglutinin by amantadine. EMBO J 9, 3469–3476.
    [Google Scholar]
  36. Taylor, G., Stott, E. J., Furze, J., Ford, J. & Sopp, P. ( 1992; ). Protective epitopes on the fusion protein of respiratory syncytial virus recognized by murine and bovine monoclonal antibodies. J Gen Virol 73, 2217–2223.[CrossRef]
    [Google Scholar]
  37. Techaarpornkul, S., Barretto, N. & Peeples, M. E. ( 2001; ). Functional analysis of recombinant respiratory syncytial virus deletion mutants lacking the small hydrophobic and/or attachment glycoprotein gene. J Virol 75, 6825–6834.[CrossRef]
    [Google Scholar]
  38. van Meer, G. & Simons, K. ( 1988; ). Lipid polarity and sorting in epithelial cells. J Cell Biochem 36, 51–58.[CrossRef]
    [Google Scholar]
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