1887

Abstract

The nucleocapsid (N) protein of hantaviruses (family ) is the most abundant component of the virion; it encapsidates genomic RNA segments and participates in viral genome transcription and replication, as well as in virus assembly. During RNA encapsidation, the N protein forms intermediate trimers and then oligomers via ‘head-to-head, tail-to-tail’ interactions. In previous work, using Tula hantavirus (TULV) N protein as a model, it was demonstrated that an intact coiled-coil structure of the N terminus is crucial for the oligomerization capacity of the N protein and that the hydrophobic ‘a’ residues from the second -helix are especially important. Here, the importance of charged amino acid residues located within the coiled-coil for trimer formation and oligomerization was analysed. To predict the interacting surfaces of the monomers, the previous model of TULV coiled-coils was first upgraded, taking advantage of the recently published crystal structure of the N-terminal coiled-coil of the Sin Nombre virus N protein. The results obtained using a mammalian two-hybrid assay suggested that conserved, charged amino acid residues within the coiled-coil make a substantial contribution to N protein oligomerization. This contribution probably involves (i) the formation of interacting surfaces of the N monomers (residues D35 and D38, located at the tip of the coiled-coil loop, and R63 appear particularly important) and (ii) stabilization of the coiled-coil via intramolecular ionic bridging (with E55 as a key player). It is hypothesized that the tips of the coiled-coils are the first to come into direct contact and thus to initiate tight packing of the three structures.

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2008-09-01
2019-11-17
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References

  1. Albertini, A. A., Schoehn, G., Weissenhorn, W. & Ruigrok, R. W. ( 2008; ). Structural aspects of rabies virus replication. Cell Mol Life Sci 65, 282–294.[CrossRef]
    [Google Scholar]
  2. Alfadhli, A., Love, Z., Arvidson, B., Seeds, J., Willey, J. & Barklis, E. ( 2001; ). Hantavirus nucleocapsid protein oligomerization. J Virol 75, 2019–2023.[CrossRef]
    [Google Scholar]
  3. Alfadhli, A., Steel, E., Finlay, L., Bachinger, H. P. & Barklis, E. ( 2002; ). Hantavirus nucleocapsid protein coiled-coil domains. J Biol Chem 277, 27103–27108.[CrossRef]
    [Google Scholar]
  4. Alminaite, A., Halttunen, V., Kumar, V., Vaheri, A., Holm, L. & Plyusnin, A. ( 2006; ). Oligomerization of hantavirus nucleocapsid protein: analysis of the N-terminal coiled-coil domain. J Virol 80, 9073–9081.[CrossRef]
    [Google Scholar]
  5. Boudko, S. P., Kuhn, R. J. & Rossmann, M. G. ( 2007; ). The coiled-coil domain structure of the Sin Nombre virus nucleocapsid protein. J Mol Biol 366, 1538–1544.[CrossRef]
    [Google Scholar]
  6. Bullough, P. A., Hughson, F. M., Skehel, J. J. & Wiley, D. C. ( 1994; ). Structure of influenza haemagglutinin at the pH of membrane fusion. Nature 371, 37–43.[CrossRef]
    [Google Scholar]
  7. Burkhard, P., Stetefeld, J. & Strelkov, S. V. ( 2001; ). Coiled coils: a highly versatile protein folding motif. Trends Cell Biol 11, 82–88.[CrossRef]
    [Google Scholar]
  8. Chen, J., Skehe, J. J. & Wiley, D. C. ( 1999; ). N- and C-terminal residues combine in the fusion-pH influenza hemagglutinin HA2 subunit to form an N cap that terminates the triple-stranded coiled coil. Proc Natl Acad Sci U S A 96, 8967–8972.[CrossRef]
    [Google Scholar]
  9. Comeau, S. R., Gatchell, D. W., Vajda, S. & Camacho, C. J. ( 2004; ). ClusPro: an automated docking and discrimination method for the prediction of protein complexes. Bioinformatics 20, 45–50.[CrossRef]
    [Google Scholar]
  10. DiCarlo, A., Möller, P., Lander, A., Kolesnikova, L. & Becker, S. ( 2007; ). Nucleocapsid formation and RNA synthesis of Marburg virus is dependent on two coiled coil motifs in the nucleoprotein. Virol J 4, 105 [CrossRef]
    [Google Scholar]
  11. Hale, B. G., Batty, I. H., Downes, C. P. & Randall, R. E. ( 2008; ). Binding of influenza A virus NS1 protein to the inter-SH2 domain of p85 suggests a novel mechanism for phosphoinositide 3-kinase activation. J Biol Chem 283, 1372–1380.[CrossRef]
    [Google Scholar]
  12. Holm, L. & Sander, C. ( 1992; ). Evaluation of protein models by atomic solvation preference. J Mol Biol 225, 93–105.[CrossRef]
    [Google Scholar]
  13. Jääskeläinen, K. M., Kaukinen, P., Minskaya, E. S., Plyusnina, A., Vapalahti, O., Elliott, R. M., Weber, F., Vaheri, A. & Plyusnin, A. ( 2007; ). Tula and Puumala hantavirus NSs ORFs are functional and the products inhibit activation of the interferon-beta promoter. J Med Virol 79, 1527–1536.[CrossRef]
    [Google Scholar]
  14. Kaukinen, P., Koistinen, V., Vapalahti, O., Vaheri, A. & Plyusnin, A. ( 2001; ). Interaction between molecules of hantavirus nucleocapsid protein. J Gen Virol 82, 1845–1853.
    [Google Scholar]
  15. Kaukinen, P., Vaheri, A. & Plyusnin, A. ( 2003a; ). Non-covalent interaction between nucleocapsid protein of Tula hantavirus and small ubiquitin-related modifier-1, SUMO-1. Virus Res 92, 37–45.[CrossRef]
    [Google Scholar]
  16. Kaukinen, P., Vaheri, A. & Plyusnin, A. ( 2003b; ). Mapping of the regions involved in homotypic interactions of Tula hantavirus N protein. J Virol 77, 10910–10916.[CrossRef]
    [Google Scholar]
  17. Kaukinen, P., Kumar, V., Tulimaki, K., Engelhardt, P., Vaheri, A. & Plyusnin, A. ( 2004; ). Oligomerization of hantavirus N protein: C-terminal α-helices interact to form a shared hydrophobic space. J Virol 78, 13669–13677.[CrossRef]
    [Google Scholar]
  18. Kaukinen, P., Vaheri, A. & Plyusnin, A. ( 2005; ). Hantavirus nucleocapsid protein: a multifunctional molecule with both housekeeping and ambassadorial duties. Arch Virol 150, 1693–1713.[CrossRef]
    [Google Scholar]
  19. Kukkonen, S. K. J., Vaheri, A. & Plyusnin, A. ( 2004; ). Tula hantavirus L protein is a 250 kDa perinuclear membrane-associated protein. J Gen Virol 85, 1181–1189.[CrossRef]
    [Google Scholar]
  20. Li, X. D., Makela, T. P., Guo, D., Soliymani, R., Koistinen, V., Vapalahti, O., Vaheri, A. & Lankinen, H. ( 2002; ). Hantavirus nucleocapsid protein interacts with the Fas-mediated apoptosis enhancer Daxx. J Gen Virol 83, 759–766.
    [Google Scholar]
  21. Lupas, A. ( 1996; ). Coiled coils: new structures and new functions. Trends Biochem Sci 21, 375–382.[CrossRef]
    [Google Scholar]
  22. Maeda, A., Lee, B. H., Yoshimatsu, K., Saijo, M., Kurane, I., Arikawa, J. & Morikawa, S. ( 2003; ). The intracellular association of the nucleocapsid protein (NP) of hantaan virus (HTNV) with small ubiquitin-like modifier-1 (SUMO-1) conjugating enzyme 9 (Ubc9). Virology 305, 288–297.[CrossRef]
    [Google Scholar]
  23. Mir, M. A. & Panganiban, A. T. ( 2004; ). Trimeric hantavirus nucleocapsid protein binds specifically to the viral RNA panhandle. J Virol 78, 8281–8288.[CrossRef]
    [Google Scholar]
  24. Möller, P., Pariente, N., Klenk, H. D. & Becker, S. ( 2005; ). Homo-oligomerization of Marburgvirus VP35 is essential for its function in replication and transcription. J Virol 79, 14876–14886.[CrossRef]
    [Google Scholar]
  25. Nichol, S. T., Beaty, B. J., Elliott, R. M., Goldbach, R., Plyusnin, A., Schmaljohn, C. S. & Tesh, R. B. ( 2005; ). Bunyaviridae. In Virus Taxonomy. Eighth Report of the International Committee on Taxonomy of Viruses, pp. 695–716. Edited by C. M. Fauquet, M. A. Mayo, J. Maniloff, U. Desselberger & L. A. Ball. Amsterdam: Elsevier Academic Press.
  26. Ramachandran, G. N., Ramakrishnan, C. & Sasisekharan, V. ( 1963; ). Stereochemistry of polypeptide chain configurations. J Mol Biol 7, 95–99.[CrossRef]
    [Google Scholar]
  27. Ravkov, E. V. & Compans, R. W. ( 2001; ). Hantavirus nucleocapsid protein is expressed as a membrane-associated protein in the perinuclear region. J Virol 75, 1808–1815.[CrossRef]
    [Google Scholar]
  28. Rost, B. ( 1997; ). Protein structures sustain evolutionary drift. Fold Des 2, S19–S24.[CrossRef]
    [Google Scholar]
  29. Sali, A. & Blundel, T. L. ( 1993; ). Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol 234, 779–815.[CrossRef]
    [Google Scholar]
  30. Sander, C. & Schneider, R. ( 1991; ). Database of homology-derived structures and the structural meaning of sequence alignment. Proteins 9, 56–68.[CrossRef]
    [Google Scholar]
  31. Severson, W.., Xu, X., Kuhn, M., Senutovitch, N., Thokala, M., Ferron, F., Longhi, S., Canard, B. & Jonsson, C. B. ( 2005; ). Essential amino acids of the Hantaan virus N protein in its interaction with RNA. J Virol 79, 10032–10039.[CrossRef]
    [Google Scholar]
  32. Spencer, K. A. & Hiscox, J. A. ( 2006; ). Characterisation of the RNA binding properties of the coronavirus infectious bronchitis virus nucleocapsid protein amino-terminal region. FEBS Lett 580, 5993–5998.[CrossRef]
    [Google Scholar]
  33. Staker, B. L., Hjerrild, K., Feese, M. D., Behnke, C. A., Burgin, A. B., Jr & Stewart, L. J. ( 2002; ). The mechanism of topoisomerase I poisoning by a camptothecin analog. Proc Natl Acad Sci U S A 99, 15387–15392.[CrossRef]
    [Google Scholar]
  34. van Marle, G., Luytjes, W., van der Most, R. G., van der Straaten, T. & Spaan, W. J. ( 1995; ). Regulation of coronavirus mRNA transcription. J Virol 69, 7851–7856.
    [Google Scholar]
  35. Vapalahti, O., Mustonen, J., Lundkvist, A., Henttonen, H., Plyusnin, A. & Vaheri, A. ( 2003; ). Hantavirus infections in Europe. Lancet Infect Dis 3, 653–661.[CrossRef]
    [Google Scholar]
  36. Vriend, G. ( 1990; ). A molecular modeling and drug design program. J Mol Graph 8, 52–56.[CrossRef]
    [Google Scholar]
  37. Xu, X., Severson, W., Villegas, N., Schmaljohn, C. S. & Jonsson, C. B. ( 2002; ). The RNA binding domain of the Hantaan virus N protein maps to a central, conserved region. J Virol 76, 3301–3308.[CrossRef]
    [Google Scholar]
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