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. 1995 Sep;69(9):5528–5534. doi: 10.1128/jvi.69.9.5528-5534.1995

Biological function of the low-pH, fusion-inactive conformation of rabies virus glycoprotein (G): G is transported in a fusion-inactive state-like conformation.

Y Gaudin 1, C Tuffereau 1, P Durrer 1, A Flamand 1, R W Ruigrok 1
PMCID: PMC189404  PMID: 7543584

Abstract

The glycoprotein (G) of rabies virus can assume at least three different conformations: the native (N) state detected at the viral surface above pH 7; the activated (A) hydrophobic state, which is probably involved in the first steps of the fusion process; and the fusion-inactive (I) conformation. There is a pH-dependent equilibrium between these states, the equilibrium being shifted towards the I state at low pH. It has been supposed that the transition from the N to the I state mediates membrane fusion. By using a lipid-mixing assay, we studied the kinetics of fusion and fusion inactivation for two rabies virus strains, PV and CVS. In addition, by using electron microscopy and a trypsin sensitivity assay, we analyzed the kinetics of the conformational change towards the I state for both strains. Although the PV strain fuses faster, inactivation and the conformational change of PV G occur more slowly than for the CVS strain. This suggests that the structural transition towards the I state is irrelevant to the fusion process. Immunofluorescence and immunoprecipitation experiments performed with infected cells and two different monoclonal antibodies, one specific for the N form of G and one which recognizes both the N and the I states, suggest that G is transported in an I state-like conformation through the Golgi apparatus and acquires its N structure only near or at the cell surface. We propose that the role of the I state is to avoid unspecific fusion during transport of G in the acidic Golgi vesicles.

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Selected References

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  1. Anderson R. G., Orci L. A view of acidic intracellular compartments. J Cell Biol. 1988 Mar;106(3):539–543. doi: 10.1083/jcb.106.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson R. G., Pathak R. K. Vesicles and cisternae in the trans Golgi apparatus of human fibroblasts are acidic compartments. Cell. 1985 Mar;40(3):635–643. doi: 10.1016/0092-8674(85)90212-0. [DOI] [PubMed] [Google Scholar]
  3. Ciampor F., Bayley P. M., Nermut M. V., Hirst E. M., Sugrue R. J., Hay A. J. 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. 1992 May;188(1):14–24. doi: 10.1016/0042-6822(92)90730-d. [DOI] [PubMed] [Google Scholar]
  4. Clague M. J., Schoch C., Zech L., Blumenthal R. Gating kinetics of pH-activated membrane fusion of vesicular stomatitis virus with cells: stopped-flow measurements by dequenching of octadecylrhodamine fluorescence. Biochemistry. 1990 Feb 6;29(5):1303–1308. doi: 10.1021/bi00457a028. [DOI] [PubMed] [Google Scholar]
  5. Doms R. W., Keller D. S., Helenius A., Balch W. E. Role for adenosine triphosphate in regulating the assembly and transport of vesicular stomatitis virus G protein trimers. J Cell Biol. 1987 Nov;105(5):1957–1969. doi: 10.1083/jcb.105.5.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fredericksen B. L., Whitt M. A. Vesicular stomatitis virus glycoprotein mutations that affect membrane fusion activity and abolish virus infectivity. J Virol. 1995 Mar;69(3):1435–1443. doi: 10.1128/jvi.69.3.1435-1443.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gaudin Y., Ruigrok R. W., Knossow M., Flamand A. Low-pH conformational changes of rabies virus glycoprotein and their role in membrane fusion. J Virol. 1993 Mar;67(3):1365–1372. doi: 10.1128/jvi.67.3.1365-1372.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gaudin Y., Ruigrok R. W., Tuffereau C., Knossow M., Flamand A. Rabies virus glycoprotein is a trimer. Virology. 1992 Apr;187(2):627–632. doi: 10.1016/0042-6822(92)90465-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gaudin Y., Tuffereau C., Segretain D., Knossow M., Flamand A. Reversible conformational changes and fusion activity of rabies virus glycoprotein. J Virol. 1991 Sep;65(9):4853–4859. doi: 10.1128/jvi.65.9.4853-4859.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hammond C., Helenius A. Folding of VSV G protein: sequential interaction with BiP and calnexin. Science. 1994 Oct 21;266(5184):456–458. doi: 10.1126/science.7939687. [DOI] [PubMed] [Google Scholar]
  11. Heinz F. X., Stiasny K., Püschner-Auer G., Holzmann H., Allison S. L., Mandl C. W., Kunz C. Structural changes and functional control of the tick-borne encephalitis virus glycoprotein E by the heterodimeric association with protein prM. Virology. 1994 Jan;198(1):109–117. doi: 10.1006/viro.1994.1013. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lobigs M., Garoff H. Fusion function of the Semliki Forest virus spike is activated by proteolytic cleavage of the envelope glycoprotein precursor p62. J Virol. 1990 Mar;64(3):1233–1240. doi: 10.1128/jvi.64.3.1233-1240.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Puri A., Grimaldi S., Blumenthal R. Role of viral envelope sialic acid in membrane fusion mediated by the vesicular stomatitis virus envelope glycoprotein. Biochemistry. 1992 Oct 20;31(41):10108–10113. doi: 10.1021/bi00156a034. [DOI] [PubMed] [Google Scholar]
  15. Ramalho-Santos J., Nir S., Düzgünes N., de Carvalho A. P., de Lima M. da C. A common mechanism for influenza virus fusion activity and inactivation. Biochemistry. 1993 Mar 23;32(11):2771–2779. doi: 10.1021/bi00062a006. [DOI] [PubMed] [Google Scholar]
  16. Ruigrok R. W., Martin S. R., Wharton S. A., Skehel J. J., Bayley P. M., Wiley D. C. Conformational changes in the hemagglutinin of influenza virus which accompany heat-induced fusion of virus with liposomes. Virology. 1986 Dec;155(2):484–497. doi: 10.1016/0042-6822(86)90210-2. [DOI] [PubMed] [Google Scholar]
  17. Stegmann T., Booy F. P., Wilschut J. Effects of low pH on influenza virus. Activation and inactivation of the membrane fusion capacity of the hemagglutinin. J Biol Chem. 1987 Dec 25;262(36):17744–17749. [PubMed] [Google Scholar]
  18. Stegmann T., Doms R. W., Helenius A. Protein-mediated membrane fusion. Annu Rev Biophys Biophys Chem. 1989;18:187–211. doi: 10.1146/annurev.bb.18.060189.001155. [DOI] [PubMed] [Google Scholar]
  19. Struck D. K., Hoekstra D., Pagano R. E. Use of resonance energy transfer to monitor membrane fusion. Biochemistry. 1981 Jul 7;20(14):4093–4099. doi: 10.1021/bi00517a023. [DOI] [PubMed] [Google Scholar]
  20. Sugrue R. J., Bahadur G., Zambon M. C., Hall-Smith M., Douglas A. R., Hay A. J. Specific structural alteration of the influenza haemagglutinin by amantadine. EMBO J. 1990 Nov;9(11):3469–3476. doi: 10.1002/j.1460-2075.1990.tb07555.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weber T., Paesold G., Galli C., Mischler R., Semenza G., Brunner J. Evidence for H(+)-induced insertion of influenza hemagglutinin HA2 N-terminal segment into viral membrane. J Biol Chem. 1994 Jul 15;269(28):18353–18358. [PubMed] [Google Scholar]
  22. Wharton S. A., Calder L. J., Ruigrok R. W., Skehel J. J., Steinhauer D. A., Wiley D. C. Electron microscopy of antibody complexes of influenza virus haemagglutinin in the fusion pH conformation. EMBO J. 1995 Jan 16;14(2):240–246. doi: 10.1002/j.1460-2075.1995.tb06997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wharton S. A., Skehel J. J., Wiley D. C. Studies of influenza haemagglutinin-mediated membrane fusion. Virology. 1986 Feb;149(1):27–35. doi: 10.1016/0042-6822(86)90083-8. [DOI] [PubMed] [Google Scholar]
  24. White J. M. Membrane fusion. Science. 1992 Nov 6;258(5084):917–924. doi: 10.1126/science.1439803. [DOI] [PubMed] [Google Scholar]
  25. White J., Kartenbeck J., Helenius A. Membrane fusion activity of influenza virus. EMBO J. 1982;1(2):217–222. doi: 10.1002/j.1460-2075.1982.tb01150.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Whitt M. A., Buonocore L., Prehaud C., Rose J. K. Membrane fusion activity, oligomerization, and assembly of the rabies virus glycoprotein. Virology. 1991 Dec;185(2):681–688. doi: 10.1016/0042-6822(91)90539-n. [DOI] [PubMed] [Google Scholar]
  27. Wiley D. C., Skehel J. J. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. Annu Rev Biochem. 1987;56:365–394. doi: 10.1146/annurev.bi.56.070187.002053. [DOI] [PubMed] [Google Scholar]

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