1887

Abstract

Flaviviruses are assembled to bud into the lumen of the endoplasmic reticulum (ER) and are secreted through the vesicle transport pathway, but the details of the molecular mechanism of virion assembly remain largely unknown. In this study, a highly conserved region in the prM protein was identified among flaviviruses. In the subviral particle (SP) system of tick-borne encephalitis virus (TBEV) and Japanese encephalitis virus, secretion of SPs was impaired by a mutation in the conserved region in the prM protein. Viral proteins were sparse in the Golgi complex and accumulated in the ER. Ultrastructural analysis revealed that long filamentous structures, rather than spherical SPs, were observed in the lumen of the ER as a result of the mutation. The production of infectious virions derived from infectious cDNA of TBEV was also reduced by mutations in the conserved region. Molecular modelling analysis suggested that the conserved region is important for the association of prM–envelope protein heterodimers in the formation of a spike of immature virion. These results are the first demonstration that the conserved region in the prM protein is a molecular determinant for the flavivirus assembly process.

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2012-01-01
2024-03-28
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References

  1. Allison S. L., Schalich J., Stiasny K., Mandl C. W., Kunz C., Heinz F. X. 1995a; Oligomeric rearrangement of tick-borne encephalitis virus envelope proteins induced by an acidic pH. J Virol 69:695–700[PubMed]
    [Google Scholar]
  2. Allison S. L., Stadler K., Mandl C. W., Kunz C., Heinz F. X. 1995b; Synthesis and secretion of recombinant tick-borne encephalitis virus protein E in soluble and particulate form. J Virol 69:5816–5820[PubMed]
    [Google Scholar]
  3. Allison S. L., Stiasny K., Stadler K., Mandl C. W., Heinz F. X. 1999; Mapping of functional elements in the stem-anchor region of tick-borne encephalitis virus envelope protein E. J Virol 73:5605–5612[PubMed]
    [Google Scholar]
  4. Allison S. L., Schalich J., Stiasny K., Mandl C. W., Heinz F. X. 2001; Mutational evidence for an internal fusion peptide in flavivirus envelope protein E. J Virol 75:4268–4275 [View Article][PubMed]
    [Google Scholar]
  5. Bieniasz P. D. 2006; Late budding domains and host proteins in enveloped virus release. Virology 344:55–63 [View Article][PubMed]
    [Google Scholar]
  6. Chambers T. J., Hahn C. S., Galler R., Rice C. M. 1990; Flavivirus genome organization, expression, and replication. Annu Rev Microbiol 44:649–688 [View Article][PubMed]
    [Google Scholar]
  7. DeLano W. L. 2002 The PyMol Molecular Graphics System San Carlos, CA: DeLano Scientific;
    [Google Scholar]
  8. Elshuber S., Allison S. L., Heinz F. X., Mandl C. W. 2003; Cleavage of protein prM is necessary for infection of BHK-21 cells by tick-borne encephalitis virus. J Gen Virol 84:183–191 [View Article][PubMed]
    [Google Scholar]
  9. Finn R. D., Mistry J., Tate J., Coggill P., Heger A., Pollington J. E., Gavin O. L., Gunasekaran P., Ceric G. other authors 2010; The Pfam protein families database. Nucleic Acids Res 38:Database issueD211–D222 [View Article][PubMed]
    [Google Scholar]
  10. Fonseca B. A., Pincus S., Shope R. E., Paoletti E., Mason P. W. 1994; Recombinant vaccinia viruses co-expressing dengue-1 glycoproteins prM and E induce neutralizing antibodies in mice. Vaccine 12:279–285 [View Article][PubMed]
    [Google Scholar]
  11. Gritsun T. S., Lisak V. M., Liapustin V. N., Korolev M. B., Lashkevich V. A. 1989; [Slowly-sedimenting hemagglutinin of the tick-borne encephalitis virus]. Vopr Virusol 34:449–454 in Russian [PubMed]
    [Google Scholar]
  12. Guirakhoo F., Heinz F. X., Mandl C. W., Holzmann H., Kunz C. 1991; Fusion activity of flaviviruses: comparison of mature and immature (prM-containing) tick-borne encephalitis virions. J Gen Virol 72:1323–1329 [View Article][PubMed]
    [Google Scholar]
  13. Guirakhoo F., Bolin R. A., Roehrig J. T. 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 191:921–931 [View Article][PubMed]
    [Google Scholar]
  14. Hayasaka D., Gritsun T. S., Yoshii K., Ueki T., Goto A., Mizutani T., Kariwa H., Iwasaki T., Gould E. A., Takashima I. 2004; Amino acid changes responsible for attenuation of virus neurovirulence in an infectious cDNA clone of the Oshima strain of tick-borne encephalitis virus. J Gen Virol 85:1007–1018 [View Article][PubMed]
    [Google Scholar]
  15. Heinz F., Kunz C. 1977; Concentration and purification of tick-borne encephalitis virus grown in suspensions of chick embryo cells. Acta Virol 21:301–307[PubMed]
    [Google Scholar]
  16. Heinz F. X., Stiasny K., Püschner-Auer G., Holzmann H., Allison S. L., Mandl C. W., Kunz C. 1994; Structural changes and functional control of the tick-borne encephalitis virus glycoprotein E by the heterodimeric association with protein prM. Virology 198:109–117 [View Article][PubMed]
    [Google Scholar]
  17. Kail M., Hollinshead M., Ansorge W., Pepperkok R., Frank R., Griffiths G., Vaux D. 1991; The cytoplasmic domain of alphavirus E2 glycoprotein contains a short linear recognition signal required for viral budding. EMBO J 10:2343–2351[PubMed]
    [Google Scholar]
  18. Keen J. H., Willingham M. C., Pastan I. H. 1979; Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets. Cell 16:303–312 [View Article][PubMed]
    [Google Scholar]
  19. Komoro K., Hayasaka D., Mizutani T., Kariwa H., Takashima I. 2000; Characterization of monoclonal antibodies against Hokkaido strain tick-borne encephalitis virus. Microbiol Immunol 44:533–536[PubMed] [CrossRef]
    [Google Scholar]
  20. Konishi E., Mason P. W. 1993; Proper maturation of the Japanese encephalitis virus envelope glycoprotein requires cosynthesis with the premembrane protein. J Virol 67:1672–1675[PubMed]
    [Google Scholar]
  21. Konishi E., Yamaoka M., Khin-Sane-Win, Kurane I., Mason P. W. 1998; Induction of protective immunity against Japanese encephalitis in mice by immunization with a plasmid encoding Japanese encephalitis virus premembrane and envelope genes. J Virol 72:4925–4930[PubMed]
    [Google Scholar]
  22. Kuhn R. J., Zhang W., Rossmann M. G., Pletnev S. V., Corver J., Lenches E., Jones C. T., Mukhopadhyay S., Chipman P. R., Strauss E. G. 2002; Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. Cell 108:717–725 [View Article][PubMed]
    [Google Scholar]
  23. Li L., Lok S. M., Yu I. M., Zhang Y., Kuhn R. J., Chen J., Rossmann M. G. 2008; The flavivirus precursor membrane–envelope protein complex: structure and maturation. Science 319:1830–1834 [View Article][PubMed]
    [Google Scholar]
  24. Lindenbach B. D., Thiel H. J., Rice C. M. 2007; Flaviviridae: the viruses and their replication. In Fields Virology, 5th edn. pp. 1101–1152 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  25. Lobigs M., Lee E. 2004; Inefficient signalase cleavage promotes efficient nucleocapsid incorporation into budding flavivirus membranes. J Virol 78:178–186 [View Article][PubMed]
    [Google Scholar]
  26. Lorenz I. C., Allison S. L., Heinz F. X., Helenius A. 2002; Folding and dimerization of tick-borne encephalitis virus envelope proteins prM and E in the endoplasmic reticulum. J Virol 76:5480–5491 [View Article][PubMed]
    [Google Scholar]
  27. Lorenz I. C., Kartenbeck J., Mezzacasa A., Allison S. L., Heinz F. X., Helenius A. 2003; Intracellular assembly and secretion of recombinant subviral particles from tick-borne encephalitis virus. J Virol 77:4370–4382 [View Article][PubMed]
    [Google Scholar]
  28. Mackenzie J. M., Westaway E. G. 2001; Assembly and maturation of the flavivirus Kunjin virus appear to occur in the rough endoplasmic reticulum and along the secretory pathway, respectively. J Virol 75:10787–10799 [View Article][PubMed]
    [Google Scholar]
  29. Mason P. W., Pincus S., Fournier M. J., Mason T. L., Shope R. E., Paoletti E. 1991; Japanese encephalitis virus–vaccinia recombinants produce particulate forms of the structural membrane proteins and induce high levels of protection against lethal JEV infection. Virology 180:294–305 [View Article][PubMed]
    [Google Scholar]
  30. Morita E., Sundquist W. I. 2004; Retrovirus budding. Annu Rev Cell Dev Biol 20:395–425 [View Article][PubMed]
    [Google Scholar]
  31. Op De Beeck A., Molenkamp R., Caron M., Ben Younes A., Bredenbeek P., Dubuisson J. 2003; Role of the transmembrane domains of prM and E proteins in the formation of yellow fever virus envelope. J Virol 77:813–820 [View Article][PubMed]
    [Google Scholar]
  32. Op De Beeck A., Rouillé Y., Caron M., Duvet S., Dubuisson J. 2004; The transmembrane domains of the prM and E proteins of yellow fever virus are endoplasmic reticulum localization signals. J Virol 78:12591–12602 [View Article][PubMed]
    [Google Scholar]
  33. Pryor M. J., Azzola L., Wright P. J., Davidson A. D. 2004; Histidine 39 in the dengue virus type 2 M protein has an important role in virus assembly. J Gen Virol 85:3627–3636 [View Article][PubMed]
    [Google Scholar]
  34. Stadler K., Allison S. L., Schalich J., Heinz F. X. 1997; Proteolytic activation of tick-borne encephalitis virus by furin. J Virol 71:8475–8481[PubMed]
    [Google Scholar]
  35. Takashima I., Morita K., Chiba M., Hayasaka D., Sato T., Takezawa C., Igarashi A., Kariwa H., Yoshimatsu K. other authors 1997; A case of tick-borne encephalitis in Japan and isolation of the virus. J Clin Microbiol 35:1943–1947[PubMed]
    [Google Scholar]
  36. Tan T. T., Bhuvanakantham R., Li J., Howe J., Ng M. L. 2009; Tyrosine 78 of premembrane protein is essential for assembly of West Nile virus. J Gen Virol 90:1081–1092 [View Article][PubMed]
    [Google Scholar]
  37. 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 63:2521–2526[PubMed]
    [Google Scholar]
  38. Whitt M. A., Chong L., Rose J. K. 1989; Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant. J Virol 63:3569–3578[PubMed]
    [Google Scholar]
  39. Wieland F., Harter C. 1999; Mechanisms of vesicle formation: insights from the COP system. Curr Opin Cell Biol 11:440–446 [View Article][PubMed]
    [Google Scholar]
  40. Yamshchikov V. F., Compans R. W. 1993; Regulation of the late events in flavivirus protein processing and maturation. Virology 192:38–51 [View Article][PubMed]
    [Google Scholar]
  41. Yoshii K., Hayasaka D., Goto A., Obara M., Araki K., Yoshimatsu K., Arikawa J., Ivanov L., Mizutani T. other authors 2003; Enzyme-linked immunosorbent assay using recombinant antigens expressed in mammalian cells for serodiagnosis of tick-borne encephalitis. J Virol Methods 108:171–179 [View Article][PubMed]
    [Google Scholar]
  42. Yoshii K., Konno A., Goto A., Nio J., Obara M., Ueki T., Hayasaka D., Mizutani T., Kariwa H., Takashima I. 2004; Single point mutation in tick-borne encephalitis virus prM protein induces a reduction of virus particle secretion. J Gen Virol 85:3049–3058 [View Article][PubMed]
    [Google Scholar]
  43. Zhang Y., Corver J., Chipman P. R., Zhang W., Pletnev S. V., Sedlak D., Baker T. S., Strauss J. H., Kuhn R. J., Rossmann M. G. 2003; Structures of immature flavivirus particles. EMBO J 22:2604–2613 [View Article][PubMed]
    [Google Scholar]
  44. Zhao H., Lindqvist B., Garoff H., von Bonsdorff C. H., Liljeström P. 1994; A tyrosine-based motif in the cytoplasmic domain of the alphavirus envelope protein is essential for budding. EMBO J 13:4204–4211[PubMed]
    [Google Scholar]
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