1887

Abstract

Intermolecular interactions of Tula hantavirus N (nucleocapsid) protein were detected in the yeast two-hybrid system, prompting further attempts to study this phenomenon. Using chemical cross-linking and immunoblotting it was shown that the N protein from purified virus and from infected cell lysates as well as recombinant protein produced in a baculovirus expression system are capable of forming dimers, trimers and multimers, thus confirming the capacity of the protein molecules to interact with each other. An ELISA format was developed in which molecules of the recombinant N protein were shown to associate non-covalently, via electrostatic interactions. Divalent cations (Ca, Mn, Mg, Ba) enhanced the process 3- to 8-fold suggesting that adequate folding of the N protein is crucial for the association. Based on these data a model for hantavirus nucleocapsid assembly is proposed, in which N molecules first trimerize around the viral RNA molecule, and then the trimers gradually assemble forming longer multimers.

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2001-08-01
2020-01-28
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References

  1. Alfadhli, A., Love, Z., Arvidson, B, Seeds, J., Willey, J. & Mackow, E. ( 2001; ). Hantavirus nucleocapsid protein oligomerization. Journal of Virology 75, 2019-2023.[CrossRef]
    [Google Scholar]
  2. Becker, S., Rinne, C., Hofsäss, U., Klenk, H.-D. & Mühlberger, E. ( 1998; ). Interactions of Marburg virus nucleocapsid proteins. Virology 249, 406-417.[CrossRef]
    [Google Scholar]
  3. Elliott, R. M., Bouloy, M., Calisher, C. H., Goldbach, R., Moyer, J. T., Nichol, S. T., Pettersson, R., Plyusnin, A. & Schmaljohn, C. S. ( 2000; ). Family Bunyaviridae. In Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses , pp. 599-621. Edited by M. H. V. van Regenmortel, C. M. Fauquet, D. H. L. Bishop, E. B. Carstens, M. K. Estes, S. M. Lemon, J. Maniloff, M. A. Mayo, D. J. McGeoch, C. R. Pringle & R. B. Wickner. San Diego:Academic Press.
  4. Fields, S. & Song, O.-K. ( 1989; ). A novel genetic system to detect protein–protein interactions. Nature 340, 245-246.[CrossRef]
    [Google Scholar]
  5. Gavrilovskaya, I. N., Shepley, M., Shaw, R., Ginsberg, M. H. & Mackow, E. R. ( 1998; ). Integrins mediate the cellular entry of hantaviruses that cause respiratory failure. Proceedings of the National Academy of Sciences, USA 95, 7074-7079.[CrossRef]
    [Google Scholar]
  6. Gavrilovskaya, I. N., Brown, E. J., Ginsberg, M. H. & Mackow, E. R. ( 1999; ). Cellular entry of hantaviruses which cause hemorrhagic fever with renal syndrome is mediated by β3 integrins. Journal of Virology 73, 3951-3959.
    [Google Scholar]
  7. Gietz, D., St Jean, A., Woods, R. A. & Schiestl, R. H. ( 1992; ). Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Research 20, 1425.[CrossRef]
    [Google Scholar]
  8. Goldsmith, C. S., Elliot, L. H., Peters, C. J. & Zaki, S. R. ( 1995; ). Ultrastructural characteristics of Sin Nombre virus, causative agent of hantavirus pulmonary syndrome. Archives of Virology 140, 2107-2122.[CrossRef]
    [Google Scholar]
  9. Gött, P., Stohwasser, R., Schnitzler, P., Darai, G. & Bautz, E. K. F. ( 1993; ). RNA binding of recombinant nucleocapsid proteins of hantaviruses. Virology 194, 332-337.[CrossRef]
    [Google Scholar]
  10. Green, L. M. & Berg, J. M. ( 1990; ). Retroviral nucleocapsid protein–metal ion intercations: folding and sequence variants. Proceedings of the National Academy of Sciences, USA 87, 6403-6407.[CrossRef]
    [Google Scholar]
  11. Handler, C. G., Eisenberg, R. J. & Cohen, G. H. ( 1996; ). Oligomeric structure of glycoproteins in herpes simplex virus type 1. Journal of Virology 70, 6067-6070.
    [Google Scholar]
  12. Haynes, J. I., Chang, D. & Consigli, R. A. ( 1993; ). Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly. Journal of Virology 67, 2486-2495.
    [Google Scholar]
  13. Krishnamurthy, S. & Samal, S. K. ( 1998; ). Identification of regions of bovine respiratory syncytial virus N protein required for binding to P protein and self-assembly. Journal of General Virology 79, 1399-1403.
    [Google Scholar]
  14. Kukkonen, S. K. J., Vaheri, A. & Plyusnin, A. ( 1998; ). Completion of the Tula hantavirus genome sequence: properties of the L segment and heterogeneity found in the 3′ termini of S and L genome RNAs. Journal of General Virology 79, 2615-2622.
    [Google Scholar]
  15. Lundkvist, Å., Fatouros, A. & Niklasson, B. ( 1991; ). Antigenic variation of European haemorrhagic fever with renal syndrome virus strains characterized using bank vole monoclonal antibodies. Journal of General Virology 72, 2097-2103.[CrossRef]
    [Google Scholar]
  16. Lundkvist, Å., Vapalahti, O., Plyusnin, A., Sjölander, K. B., Niklasson, B. & Vaheri, A. ( 1996; ). Characterization of Tula virus antigenic determinants defined by monoclonal antibodies raised against baculovirus expressed nucleocapsid protein. Virus Research 45, 29-44.[CrossRef]
    [Google Scholar]
  17. McInerney, T. L., El Ahmar, W., Kemp, B. E. & Poumbourios, P. ( 1998; ). Mutation-directed chemical cross-linking of human immunodeficiency virus type 1 gp41 oligomer. Journal of Virology 72, 1523-1533.
    [Google Scholar]
  18. Matsumoto, M., Hwang, S. B., Jeng, K.-S., Zhu, N. & Lai, M. M. C. ( 1996; ). Homotypic interaction and multimerization of hepatitis C virus core protein. Virology 218, 43-51.[CrossRef]
    [Google Scholar]
  19. Myers, T. M., Pieters, A. & Moyer, S. A. ( 1997; ). A highly conserved region of the Sendai virus nucleocapsid protein contributes to the NP-NP binding domain. Virology 229, 322-335.[CrossRef]
    [Google Scholar]
  20. Nemirov, K., Vapalahti, O., Lundkvist, Å., Vasilenko, V., Golovljova, I., Plyusnina, A., Niemimaa, J., Laakkonen, J., Henttonen, H., Vaheri, A. & Plyusnin, A. ( 1999; ). Isolation and characterization of Dobrava hantavirus carried by the striped field mouse (Apodemus agrarius) in Estonia. Journal of General Virology 80, 371-379.
    [Google Scholar]
  21. Paintsil, J., Muller, M., Picken, M., Gissman, L. & Zhou, J. ( 1998; ). Calcium is required in reassembly of bovine papillomavirus in vitro. Journal of General Virology 79, 1133-1141.
    [Google Scholar]
  22. Piiparinen, H., Vapalahti, O., Plyusnin, A., Vaheri, A. & Lankinen, H. ( 1997; ). Sequence analysis of the Puumala hantavirus Sotkamo strain L segment. Virus Research 51, 1-7.[CrossRef]
    [Google Scholar]
  23. Plyusnin, A., Vapalahti, O., Lankinen, H., Lehväslaiho, H., Apekina, N., Myasnikov, Y., Kallio-Kokko, H., Henttonen, H., Lundkvist, Å., Brummer-Korvenkontio, M., Gavrilovskaya, I. & Vaheri, A. ( 1994; ). Tula virus: a newly detected hantavirus carried by European common voles. Journal of Virology 68, 7833-7839.
    [Google Scholar]
  24. Plyusnin, A., Vapalahti, O. & Vaheri, A. ( 1996; ). Hantaviruses: genome structure, expression and evolution. Journal of General Virology 77, 2677-2687.[CrossRef]
    [Google Scholar]
  25. Ravkov, E. V., Nichol, S. T. & Compans, R. W. ( 1997; ). Polarized entry and release in epithelial cells of Black Creek Canal virus, a New World hantavirus. Journal of Virology 71, 1147-1154.
    [Google Scholar]
  26. Ravkov, E. V., Nichol, S. T., Peters, C. J. & Compans, R. W. ( 1998; ). Role of actin microfilaments in Black Creek Canal virus morphogenesis. Journal of Virology 72, 2865-2870.
    [Google Scholar]
  27. Ravkov, E. V. & Compans, R. W. ( 2001; ). Hantavirus nucleocapsid protein is expressed as a membrane-associated protein in the perinuclear region. Journal of Virology 75, 1808-1815.[CrossRef]
    [Google Scholar]
  28. Rodgers, R. E. D. & Consigli, R. A. ( 1996; ). Characterization of a calcium binding domain in the VP1 protein of the avian polyomavirus, budgerigar fledgling disease virus. Virus Research 44, 123-135.[CrossRef]
    [Google Scholar]
  29. Sandalon, Z. & Oppenheim, A. ( 1997; ). Self-assembly and protein-protein interactions between the SV40 capsid proteins produced in insect cells. Virology 237, 414-421.[CrossRef]
    [Google Scholar]
  30. Severson, W., Partin, L., Schmaljohn, C. S. & Jonsson, C. B. ( 1999; ). Characterization of the Hantaan nucleocapsid protein–ribonucleic acid interaction. Journal of Biological Chemistry 274, 33732-33739.[CrossRef]
    [Google Scholar]
  31. Vapalahti, O., Kallio-Kokko, H., Salonen, E.-M., Brummer-Korvenkontio, M. & Vaheri, A. ( 1992; ). Cloning and sequencing of Puumala virus Sotkamo strain S and M RNA segments: evidence for strain variation in hantaviruses and expression of the nucleocapsid protein. Journal of General Virology 73, 829-838.[CrossRef]
    [Google Scholar]
  32. Vapalahti, O., Kallio-Kokko, H., Närvänen, A., Julkunen, I., Lundkvist, A., Plyusnin, A., Lehväslaiho, H., Brummer-Korvenkontio, M., Vaheri, A. & Lankinen, H. ( 1995; ). Human B-cell epitopes of Puumala virus nucleocapsid protein, the major antigen in early serological response. Journal of Medical Virology 46, 293-303.[CrossRef]
    [Google Scholar]
  33. Vapalahti, O., Lundkvist, A., Kukkonen, S. K., Cheng, Y., Gilljam, M., Kanerva, M., Manni, T., Pejcoch, M., Niemimaa, J., Kaikusalo, A., Henttonen, H., Vaheri, A. & Plyusnin, A. ( 1996a; ). Isolation and characterization of Tula virus, a distinct serotype in the genus Hantavirus, family Bunyaviridae. Journal of General Virology 77, 3063-3067.[CrossRef]
    [Google Scholar]
  34. Vapalahti, O., Lundkvist, Å., Kallio-Kokko, H., Paukku, K., Julkunen, I., Lankinen, H. & Vaheri, A. ( 1996b; ). Antigenic properties and diagnostic potential of Puumala virus nucleocapsid protein expressed in insect cells. Journal of Clinical Microbiology 34, 119-125.
    [Google Scholar]
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