1887

Abstract

We have previously shown that respiratory syncytial virus (RSV) assembly occurs within regions of the host-cell surface membrane that are enriched in the protein caveolin-1 (cav-1). In this report, we have employed immunofluorescence microscopy to further examine the RSV assembly process. Our results show that RSV matures at regions of the cell surface that, in addition to cav-1, are enriched in the lipid-raft ganglioside GM1. Furthermore, a comparison of mock-infected and RSV-infected cells by confocal microscopy revealed a significant change in the cellular distribution of phosphocaveolin-1 (pcav-1). In mock-infected cells, pcav-1 was located at regions of the cell that interact with the extracellular matrix, termed focal adhesions (FA). In contrast, RSV-infected cells showed both a decrease in the levels of pcav-1 associated with FA and the appearance of pcav-1-containing cytoplasmic vesicles, the latter being absent in mock-infected cells. These cytoplasmic vesicles were clearly visible between 9 and 18 h post-infection and coincided with the formation of RSV filaments, although we did not observe a direct association of pcav-1 with mature virus. In addition, we noted a strong colocalization between pcav-1 and growth hormone receptor binding protein-7 (Grb7), within these cytoplasmic vesicles, which was not observed in mock-infected cells. Collectively, these findings show that the RSV assembly process occurs within specialized lipid-raft structures on the host-cell plasma membrane, induces the cellular redistribution of pcav-1 and results in the formation of cytoplasmic vesicles that contain both pcav-1 and Grb7.

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2002-08-01
2024-03-29
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References

  1. Ali A., Nayak D. P. 2000; Assembly of Sendai virus: M protein interacts with F and HN proteins and with the cytoplasmic tail and transmembrane domain of F protein. Virology 276:289–303
    [Google Scholar]
  2. Aoki T., Nomura R., Fujimoto T. 1999; Tyrosine phosphorylation of caveolin-1 in the endothelium. Experimental Cell Research 253:629–636
    [Google Scholar]
  3. Bohn W., Rutter G., Hohenberg H., Mannweiler K., Nobis P. 1986; Involvement of actin filaments in budding of measles virus: studies on cytoskeletons of infected cells. Virology 149:91–106
    [Google Scholar]
  4. Brown D. A., London E. 1998; Functions of lipid rafts in biological membranes. Annual Review of Cell and Development Biology 14:111–136
    [Google Scholar]
  5. Brown G., Aitken J., Rixon H. W. McL., Sugrue R. J. 2002; Caveolin-1 is incorporated into mature respiratory syncytial virus particles during virus assembly on the surface of virus-infected cells. Journal of General Virology 83:611–621
    [Google Scholar]
  6. Burke E., Dupuy L., Wall C., Barik S. 1998; Role of cellular actin in the gene expression and morphogenesis of human respiratory syncytial virus. Virology 252:137–148
    [Google Scholar]
  7. Cain T. J., Lui Y. J., Takizama T., Robinson J. M. 1995; Solubilization of glycosyl-phosphatidylinositol-anchored proteins in quiescent and stimulated neutrophils. Biochimica et Biophysica Acta 1235:69–78
    [Google Scholar]
  8. Caselli A., Taddei M. L., Manao G., Camici G., Ramponi G. 2001; Tyrosine-phosphorylated caveolin is a physiological substrate of the low Mr protein-tyrosine phosphatase. Journal of Biological Chemistry 276:18849–18854
    [Google Scholar]
  9. Cudmore S., Cossart P., Griffiths G., Way M. 1995; Actin-based motility of vaccinia virus. Nature 378:636–638
    [Google Scholar]
  10. Damsky C. H., Sheffield J. B., Tuszynski G. P., Warren L. 1977; Is there a role for actin in virus budding?. Journal of Cell Biology 75:593–605
    [Google Scholar]
  11. Engelman J. A., Wykoff C. C., Yasuhara S., Song K. S., Okamoto T., Lisanti M. P. 1997; Recombinant expression of caveolin-1 in oncogenically transformed cells abrogates anchorage-independent growth. Journal of Biological Chemistry 272:16374–16381
    [Google Scholar]
  12. Fujimoto T., Miyawaki A., Mikoshiba K. 1995; Inositol 1,4,5-trisphosphate receptor-like protein in plasmalemmal caveolae is linked to actin filaments. Journal of Cell Science 108:7–15
    [Google Scholar]
  13. Glenney J. R., Zokas L. 1989; Novel tyrosine kinase substrates from Rous sarcoma virus transformed cells are present in the membrane cytoskeleton. Journal of Cell Biology 108:2401–2408
    [Google Scholar]
  14. Gower T. L., Peeples M. E., Collins P. L., Graham B. S. 2001; RhoA is activated during respiratory syncytial virus infection. Virology 283:188–196
    [Google Scholar]
  15. Han D. C., Shen T. L., Guan J. L. 2001; The Grb7 family proteins: structure, interactions with other signalling molecules and potential cellular functions. Oncogene 20:6315–6321
    [Google Scholar]
  16. Harder T., Scheiffele P., Verkade P., Simons K. 1998; Lipid domain structure of the plasma membrane revealed by patching of membrane components. Journal of Cell Biology 141:929
    [Google Scholar]
  17. Kurzchalia T. V., Dupree P., Monier S. 1994; VIP21-caveolin, a protein of the trans-Golgi network and caveolae. FEBS Letters 346:88–91
    [Google Scholar]
  18. Lee H., Volonte D., Galbiati F., Iyengar P., Lublin D. M., Bregman D. B., Wilson M. T., Campos-Gonzalez R., Bouzahzah B., Pestell R. G., Scherer P. E., Lisanti M. P. 2000; Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo: identification of a c-Src/Cav-1/Grb7 signaling cassette. Molecular Endocrinology 14:1750–1775
    [Google Scholar]
  19. Manie S. N., Debreyne S., Vincent S., Gerlier D. 2000; Measles virus structural components are enriched into lipid raft microdomains: a potential cellular location for virus assembly. Journal of Virology 74:305–311
    [Google Scholar]
  20. Nguyen D. H., Hildreth J. E. 2000; Evidence for budding of human immunodeficiency virus type 1 selectively from glycolipid-enriched membrane lipid rafts. Journal of Virology 74:3264–3272
    [Google Scholar]
  21. Nomura R., Fujimoto T. 1999; Tyrosine-phosphorylated caveolin-1: immunolocalization and molecular characterization. Molecular Biology of the Cell 10:975–986
    [Google Scholar]
  22. Parry J. E., Shirodaria P. V., Pringle C. R. 1979; Pneumoviruses: the cell surface of lytically and persistently infected cells. Journal of General Virology 44:479–491
    [Google Scholar]
  23. Parton R. G. 1994; Ultrastructural localization of gangliosides; GM1 is concentrated in caveolae. Journal of Histochemistry and Cytochemistry 42:155–166
    [Google Scholar]
  24. Parton R. G., Way M., Zzorzi N., Stang E. 1997; Caveolin-3 associates with developing T-tubules during muscle differentiation. Journal of Cell Biology 136:137–154
    [Google Scholar]
  25. Pawlak G., Helfman D. M. 2001; Cytoskeletal changes in cell transformation and tumorigenesis. Current Opinions in Genetics and Development 11:41–47
    [Google Scholar]
  26. Pickl W. F., Pimentel-Muinos F. X., Seed B. 2001; Lipid rafts and pseudotyping. Journal of Virology 75:7175–7183
    [Google Scholar]
  27. Rey O., Canon J., Krogstad P. 1996; HIV-1 Gag protein associates with F-actin present in microfilaments. Virology 220:530–534
    [Google Scholar]
  28. Roberts S. R., Compans R. W., Wertz G. W. 1995; Respiratory syncytial virus matures at the apical surfaces of polarized epithelial cells. Journal of Virology 69:2667–2673
    [Google Scholar]
  29. Rothberg K. G., Heuser J. E., Donzell W. C., Ying Y. S., Glenney J. R., Anderson R. G. 1992; Caveolin, a protein component of caveolae membrane coats. Cell 68:673–682
    [Google Scholar]
  30. Sasaki H., Nakamura M., Ohno T., Matsuda Y., Yuda Y., Nonomura Y. 1995; Myosin-actin interaction plays an important role in human immunodeficiency virus type 1 release from host cells. Proceedings of the National Academy of Sciences, USA 92:2026–2030
    [Google Scholar]
  31. Scheiffele P., Verkade P., Fra A. M., Virta H., Simons K., Ikonen E. 1998; Caveolin-1 and -2 in the exocytic pathway of MDCK cells. Journal of Cell Biology 140:795–806
    [Google Scholar]
  32. Scheiffele P., Rietveld A., Wilk T., Simons K. 1999; Influenza viruses select ordered lipid domains during budding from the plasma membrane. Journal of Biological Chemistry 274:2038–2044
    [Google Scholar]
  33. Schlegel A., Arvan P., Lisanti M. P. 2001; Caveolin-1 binding to endoplasmic reticulum membranes and entry into the regulated secretory pathway are regulated by serine phosphorylation. . Journal of Biological Chemistry 276:4398–4408
    [Google Scholar]
  34. Smart E. J., Foster D. C., Ying Y. S., Kaman B. A., Anderson R. G. 1994; Protein kinase C activators inhibit receptor-mediated potocytosis by preventing internalization of caveolae. Journal of Cell Biology 124:307–313
    [Google Scholar]
  35. Smart E. J., Graf G. A., McNiven M. A., Sessa W. C., Engleman J. A., Scherer P. E., Okamoto T., Lisanti M. P. 1999; Caveolins, liquid-ordered domains, and signal transduction. Molecular and Cell Biology 19:7289–7304
    [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. Journal of General Virology 73:2217–2223
    [Google Scholar]
  37. Ulloa L., Serra R., Asenjo A., Villanueva N. 1998; Interactions between cellular actin and human respiratory syncytial virus (HRSV). Virus Research 53:13–25
    [Google Scholar]
  38. Vincent S., Gerlier D., Manie S. N. 2000; Measles virus assembly within membrane rafts. Journal of Virology 74:9911–9915
    [Google Scholar]
  39. Volonte D., Galbiati F., Pestell R. G., Lisanti M. P. 2001; Cellular stress induces the tyrosine phosphorylation of caveolin-1 (Tyr14) via activation of p38 mitogen-activated protein kinase and c-Src kinase. Evidence for caveolae, the actin cytoskeleton, and focal adhesions as mechanical sensors of osmotic stress. Journal of Biological Chemistry 276:8094–8103
    [Google Scholar]
  40. Werling D., Hope J. C., Chaplin P., Collins R. A., Taylor G., Howard C. J. 1999; Involvement of caveolae in the uptake of respiratory syncytial virus antigen by dendritic cells. Journal of Leukocyte Biology 66:50–58
    [Google Scholar]
  41. Yamada E. 1955; The fine structure of the gall bladder epithelium of the mouse. Journal of Biophysics, Biochemistry and Cytology 1:445–458
    [Google Scholar]
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