The interactions of the flavivirus envelope proteins: implications for virus entry and release

  • F. X. Heinz
  • G. Auer
  • K. Stiasny
  • H. Holzmann
  • C. Mandl
  • F. Guirakhoo
  • C. Kunz
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 9)


Viral membrane proteins play an important role in the assembly and disassembly of enveloped viruses. Oligomerization and proteolytic cleavage events are involved in controlling the functions of these proteins during virus entry and release. Using tick-borne encephalitis virus as a model we have studied the role of the flavivirus envelope proteins E and prM/M in these processes. Experiments with acidotropic agents provide evidence that the virus is taken up by receptor-mediated endocytosis and that the acidic pH in endosomes plays an important role for virus entry. The envelope glycoprotein E undergoes irreversible conformational changes at acidic pH, as indicated by the loss of several monoclonal antibody-defined epitopes, which coincide with the viral fusion activity in vitro. Sedimentation analysis reveals that these conformational changes lead to aggregation of virus particles, apparently by the exposure of hydrophobic sequence elements. None of these features are exhibited by immature virions containing E and prM rather than E and M. Detergent solubilization, sedimentation, and crosslinking experiments provide evidence that prM forms a complex with protein E which prevents the conformational changes necessary for fusion activity. The functional role of prM before its endoproteolytic cleavage by a cellular protease thus seems to be the protection of protein E from acid- inactivation during its passage through acidic trans Golgi vesicles in the course of virus release.


West Nile Virus Virus Entry Semliki Forest Virus Fusion Activity Viral Envelope Protein 
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  1. 1.
    Anderson RGW, Orci L (1988) A review of acidic intracellular compartments. J Cell Biol 106: 539–543PubMedCrossRefGoogle Scholar
  2. 2.
    Chambers TJ, Hahn CS, Galler R, Rice CM (1990) Flavivirus genome organiza¬tion, expression, and replication. Annu Rev Microbiol 44: 649–688PubMedCrossRefGoogle Scholar
  3. 3.
    Ciampor F, Bayley PM, Nermut MV, Hirst EMA, Sugrue RJ, Hay AJ (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: 14–24PubMedCrossRefGoogle Scholar
  4. 4.
    Gollins SW, Porterfield JS (1986) pH-dependent fusion between the flavivirus West Nile and liposomal model membranes. J Gen Virol 67: 157–166PubMedCrossRefGoogle Scholar
  5. 5.
    Guirakhoo F, Heinz FX, Mandl CW, Holzmann H, Kunz C (1991) Fusion activity of flaviviruses: comparison of mature and immature (prM-containing) tick-borne encephalitis virions. J Gen Virol 72: 1323–1329PubMedCrossRefGoogle Scholar
  6. 6.
    Guirakhoo F, Bolin RA, Roehrig JT (1992) The Murray Valley encephalitis virus prM protein confers acid resistance to virus particles and alters the expression of epitopes within the R2 domain of E glycoprotein. Virology 191: 921–931PubMedCrossRefGoogle Scholar
  7. 7.
    Hase T, Summers PL, Eckels KH (1989) Flavivirus entry into cultured mosquito cells and human peripheral blood monocytes. Arch Virol 104: 129–143PubMedCrossRefGoogle Scholar
  8. 8.
    Heinz FX, Kunz C (1980) Isolation of dimeric glycoprotein subunits from tick- borne encephalitis virus. Intervirology 13: 169–177PubMedCrossRefGoogle Scholar
  9. 9.
    Heinz FX, Mandl CW, Holzmann H, Kunz C, Harris BA, Rey F, Harrison SC (1991) The flavivirus envelope protein E: isolation of a soluble form from tick- borne encephalitis virus and its crystallization. J Virol 65: 5570–5583Google Scholar
  10. 10.
    Kielian M, Jungerwirth S (1990) Mechanisms of virus entry into cells. Mol Biol Med 7: 17–31PubMedGoogle Scholar
  11. 11.
    Kimura T, Gollins SW, Porterfield JS (1986) The effect of pH on the early interaction of West Nile virus with P 388 D1 cells. J Gen Virol 67: 2423–2433PubMedCrossRefGoogle Scholar
  12. 12.
    Mandl CW, Guirakhoo F, Holzmann H, Heinz FX, Kunz C (1989) Antigenic structure of the flavivirus envelope protein E at the molecular level, using tick- borne encephalitis as a model. J Virol 63: 564–571PubMedGoogle Scholar
  13. 13.
    Marsh M, Helenius A (1989) Virus entry into animal cells. In: Maramorosch K, Murphy FA, Shatkin A (eds) Advances in virus research, vol 36. Academic Press, San Diego, pp 107–151Google Scholar
  14. 14.
    Nowak T, Wengler G (1987) Analysis of disulfides present in the membrane proteins of the West Nile flavivirus. Virology 156: 127–137PubMedCrossRefGoogle Scholar
  15. 15.
    Pinto LH, Holsinger LJ, Lamb RA (1992) Influenza virus M2 protein has ion channel activity. Cell 69: 517–528PubMedCrossRefGoogle Scholar
  16. 16.
    Randolph VB, Winkler G, Stollar V (1990) Acidotropic amines inhibit proteolytic processing of flavivirus prM protein. Virology 174: 450–458PubMedCrossRefGoogle Scholar
  17. 17.
    Roehrig JT, Hunt A, Johnson AJ, Hawkes RA (1989) Synthetic peptides derived from the deduced amino acid sequence of the E-glycoprotein of Murray Valley encephalitis virus elicit antiviral antibody. Virology 171: 49–60PubMedCrossRefGoogle Scholar
  18. 18.
    Roehrig JT, Johnson AJ, Hunt AR, Bolin RA, Chu MC (1990) Antibodies to dengue 2 virus E-glycoprotein synthetic peptides identify antigenic conformation. Virology 177: 668–675PubMedCrossRefGoogle Scholar
  19. 19.
    Strauss JH, Strauss EG, Hahn CS, Hahn YS, Galler R, Hardy WR, Rice CM (1987) Replication of alphaviruses and flaviviruses: proteolytic processing of poly- proteins. In: Brinton MA, Rueckert RR (eds) Positive strand RNA viruses. Alan R. Liss, New York, pp 209–225Google Scholar
  20. 20.
    Wahlberg JM, Boere WAM, Garoff H (1989) The heterodimeric association between the membrane protein of Semliki Forest virus changes its sensitivity to low pH during virus maturation. J Virol 63: 4991–4997PubMedGoogle Scholar
  21. 21.
    Wengler G, Wengler G (1989) Cell-associated West Nile flavivirus is covered with E + Pre-M protein heterodimers which are destroyed and reorganized by proteolytic cleavage during virus release. J Virol 63: 2521–2526PubMedGoogle Scholar
  22. 22.
    White J (1990) Viral and cellular membrane fusion proteins. Annu Rev Physiol 52: 675–697PubMedCrossRefGoogle Scholar
  23. 23.
    Yoshimori T, Yomamoto A, Moriyami Y, Futai M, Tashiro Y (1991) Bafilomycin Al, a specific inhibitor of vacuolar-type H+-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. J Biol Chem 266:17707–17712PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • F. X. Heinz
    • 1
  • G. Auer
    • 1
  • K. Stiasny
    • 1
  • H. Holzmann
    • 1
  • C. Mandl
    • 1
  • F. Guirakhoo
    • 1
  • C. Kunz
    • 1
  1. 1.Institute of VirologyUniversity of ViennaViennaAustria

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