1887

Abstract

The flavivirus non-structural protein 1 (NS1) is a glycoprotein that is secreted as a soluble hexameric complex during the course of natural infection. Growing evidence indicates that this secreted form of NS1 (sNS1) plays a significant role in immune evasion and modulation during infection. Attempts to determine the crystal structure of NS1 have been unsuccessful to date and relatively little is known about the macromolecular organization of the sNS1 hexamer. Here, we have applied single-particle analysis to images of baculovirus-derived recombinant dengue 2 virus NS1 obtained by electron microscopy to determine its 3D structure to a resolution of 23 Å. This structure reveals a barrel-like organization of the three dimeric units that comprise the hexamer and provides further insights into the overall organization of oligomeric sNS1.

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2012-04-01
2021-10-24
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References

  1. Avirutnan P., Punyadee N., Noisakran S., Komoltri C., Thiemmeca S., Auethavornanan K., Jairungsri A., Kanlaya R., Tangthawornchaikul N.other authors 2006; Vascular leakage in severe dengue virus infections: a potential role for the nonstructural viral protein NS1 and complement. J Infect Dis 193:1078–1088 [View Article][PubMed]
    [Google Scholar]
  2. Avirutnan P., Zhang L., Punyadee N., Manuyakorn A., Puttikhunt C., Kasinrerk W., Malasit P., Atkinson J. P., Diamond M. S. 2007; Secreted NS1 of dengue virus attaches to the surface of cells via interactions with heparan sulfate and chondroitin sulfate E. PLoS Pathog 3:e183 [View Article][PubMed]
    [Google Scholar]
  3. Avirutnan P., Fuchs A., Hauhart R. E., Somnuke P., Youn S., Diamond M. S., Atkinson J. P. 2010; Antagonism of the complement component C4 by flavivirus nonstructural protein NS1. J Exp Med 207:793–806 [View Article][PubMed]
    [Google Scholar]
  4. Blitvich B. J., Scanlon D., Shiell B. J., Mackenzie J. S., Pham K., Hall R. A. 2001; Determination of the intramolecular disulfide bond arrangement and biochemical identification of the glycosylation sites of the nonstructural protein NS1 of Murray Valley encephalitis virus. J Gen Virol 82:2251–2256[PubMed]
    [Google Scholar]
  5. Brandt W. E., Chiewslip D., Harris D. L., Russell P. K. 1970; Partial purification and characterization of a dengue virus soluble complement-fixing antigen. J Immunol 105:1565–1568[PubMed]
    [Google Scholar]
  6. Chappell K. J., Stoermer M. J., Fairlie D. P., Young P. R. 2008; West Nile virus NS2B/NS3 protease as an antiviral target. Curr Med Chem 15:2771–2784 [View Article][PubMed]
    [Google Scholar]
  7. Chou H.-T., Luccio E., Wilson D., Stahlberg H. 2006; Cryo-electron microscopy analysis of a 105 kDa protein particle: the xylulose kinase from E.coli. Microsc Microanal 12:S02408–409 [View Article]
    [Google Scholar]
  8. Chua J. J., Bhuvanakantham R., Chow V. T., Ng M. L. 2005; Recombinant non-structural 1 (NS1) protein of dengue-2 virus interacts with human STAT3β protein. Virus Res 112:85–94 [View Article][PubMed]
    [Google Scholar]
  9. Chung K. M., Nybakken G. E., Thompson B. S., Engle M. J., Marri A., Fremont D. H., Diamond M. S. 2006a; Antibodies against West Nile virus nonstructural protein NS1 prevent lethal infection through Fc γ receptor-dependent and -independent mechanisms. J Virol 80:1340–1351 [View Article][PubMed]
    [Google Scholar]
  10. Chung K. M., Liszewski M. K., Nybakken G., Davis A. E., Townsend R. R., Fremont D. H., Atkinson J. P., Diamond M. S. 2006b; West Nile virus nonstructural protein NS1 inhibits complement activation by binding the regulatory protein factor H. Proc Natl Acad Sci U S A 103:19111–19116 [View Article][PubMed]
    [Google Scholar]
  11. Crabtree M. B., Kinney R. M., Miller B. R. 2005; Deglycosylation of the NS1 protein of dengue 2 virus, strain 16681: construction and characterization of mutant viruses. Arch Virol 150:771–786 [View Article][PubMed]
    [Google Scholar]
  12. Crooks A. J., Lee J. M., Easterbrook L. M., Timofeev A. V., Stephenson J. R. 1994; The NS1 protein of tick-borne encephalitis virus forms multimeric species upon secretion from the host cell. J Gen Virol 75:3453–3460 [View Article][PubMed]
    [Google Scholar]
  13. Deubel V., Kinney R. M., Trent D. W. 1988; Nucleotide sequence and deduced amino acid sequence of the nonstructural proteins of dengue type 2 virus, Jamaica genotype: comparative analysis of the full-length genome. Virology 165:234–244 [View Article][PubMed]
    [Google Scholar]
  14. Falconar, A. K. (1993 Dengue virus NS1. PhD thesis, University of London, London
  15. Falconar A. K., Young P. R. 1991; Production of dimer-specific and dengue virus group cross-reactive mouse monoclonal antibodies to the dengue 2 virus non-structural glycoprotein NS1. J Gen Virol 72:961–965 [View Article][PubMed]
    [Google Scholar]
  16. Falconar A. K., Young P. R., Miles M. A. 1994; Precise location of sequential dengue virus subcomplex and complex B cell epitopes on the nonstructural-1 glycoprotein. Arch Virol 137:315–326 [View Article][PubMed]
    [Google Scholar]
  17. Falgout B., Bray M., Schlesinger J. J., Lai C. J. 1990; Immunization of mice with recombinant vaccinia virus expressing authentic dengue virus nonstructural protein NS1 protects against lethal dengue virus encephalitis. J Virol 64:4356–4363[PubMed]
    [Google Scholar]
  18. Flamand M., Megret F., Mathieu M., Lepault J., Rey F. A., Deubel V. 1999; Dengue virus type 1 nonstructural glycoprotein NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion. J Virol 73:6104–6110[PubMed]
    [Google Scholar]
  19. Gutsche I., Coulibaly F., Voss J. E., Salmon J., d’Alayer J., Ermonval M., Larquet E., Charneau P., Krey T.other authors 2011; Secreted dengue virus nonstructural protein NS1 is an atypical barrel-shaped high-density lipoprotein. Proc Natl Acad Sci U S A 108:8003–8008 [View Article][PubMed]
    [Google Scholar]
  20. Hall R. A., Khromykh A. A., Mackenzie J. M., Scherret J. H., Khromykh T. I., Mackenzie J. S. 1999; Loss of dimerisation of the nonstructural protein NS1 of Kunjin virus delays viral replication and reduces virulence in mice, but still allows secretion of NS1. Virology 264:66–75 [View Article][PubMed]
    [Google Scholar]
  21. Henchal E. A., Henchal L. S., Schlesinger J. J. 1988; Synergistic interactions of anti-NS1 monoclonal antibodies protect passively immunized mice from lethal challenge with dengue 2 virus. J Gen Virol 69:2101–2107 [View Article][PubMed]
    [Google Scholar]
  22. Jawhari A., Uhring M., De Carlo S., Crucifix C., Tocchini-Valentini G., Moras D., Schultz P., Poterszman A. 2006; Structure and oligomeric state of human transcription factor TFIIE. EMBO Rep 7:500–505[PubMed]
    [Google Scholar]
  23. Krishna V. D., Rangappa M., Satchidanandam V. 2009; Virus-specific cytolytic antibodies to nonstructural protein 1 of Japanese encephalitis virus effect reduction of virus output from infected cells. J Virol 83:4766–4777 [View Article][PubMed]
    [Google Scholar]
  24. Kurosu T., Chaichana P., Yamate M., Anantapreecha S., Ikuta K. 2007; Secreted complement regulatory protein clusterin interacts with dengue virus nonstructural protein 1. Biochem Biophys Res Commun 362:1051–1056 [View Article][PubMed]
    [Google Scholar]
  25. Leblois H., Young P. R. 1995; Maturation of the dengue-2 virus NS1 protein in insect cells: effects of downstream NS2A sequences on baculovirus-expressed gene constructs. J Gen Virol 76:979–984 [View Article][PubMed]
    [Google Scholar]
  26. Lindenbach B. D., Rice C. M. 2003; Molecular biology of flaviviruses. Adv Virus Res 59:23–61 [View Article][PubMed]
    [Google Scholar]
  27. Lopez J. A., Guillen M., Sanchez-Fauquier A., Melero J. A. 1986; An antigen-binding assay to determine the specificity of monoclonal antibodies against influenza virus and mapping of epitopes. J Virol Methods 13:255–264 [View Article][PubMed]
    [Google Scholar]
  28. Ludtke S. J., Baldwin P. R., Chiu W. 1999; EMAN: semiautomated software for high-resolution single-particle reconstructions. J Struct Biol 128:82–97 [View Article][PubMed]
    [Google Scholar]
  29. Ludtke S. J., Jakana J., Song J. L., Chuang D. T., Chiu W. 2001; A 11.5 A single particle reconstruction of GroEL using EMAN. J Mol Biol 314:253–262 [View Article][PubMed]
    [Google Scholar]
  30. MacKenzie J. M., Jones M. K., Young P. R. 1996; Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication. Virology 220:232–240 [View Article][PubMed]
    [Google Scholar]
  31. Mackow E., Makino Y., Zhao B. T., Zhang Y. M., Markoff L., Buckler-White A., Guiler M., Chanock R., Lai C. J. 1987; The nucleotide sequence of dengue type 4 virus: analysis of genes coding for nonstructural proteins. Virology 159:217–228 [View Article][PubMed]
    [Google Scholar]
  32. Mandl C. W., Heinz F. X., Stöckl E., Kunz C. 1989; Genome sequence of tick-borne encephalitis virus (Western subtype) and comparative analysis of nonstructural proteins with other flaviviruses. Virology 173:291–301 [View Article][PubMed]
    [Google Scholar]
  33. Mason P. W. 1989; Maturation of Japanese encephalitis virus glycoproteins produced by infected mammalian and mosquito cells. Virology 169:354–364 [View Article][PubMed]
    [Google Scholar]
  34. Muller D. A., Young P. R. 2011; The many faces of the flavivirus non-structural glycoprotein NS1. In Molecular Virology and Control of Flaviviruses pp. 51–75 Edited by Shi P.-Y. Norfolk: Caister Academic Press;
    [Google Scholar]
  35. Noisakran S., Dechtawewat T., Avirutnan P., Kinoshita T., Siripanyaphinyo U., Puttikhunt C., Kasinrerk W., Malasit P., Sittisombut N. 2008; Association of dengue virus NS1 protein with lipid rafts. J Gen Virol 89:2492–2500 [View Article][PubMed]
    [Google Scholar]
  36. Panyasrivanit M., Khakpoor A., Wikan N., Smith D. R. 2009; Co-localization of constituents of the dengue virus translation and replication machinery with amphisomes. J Gen Virol 90:448–456 [View Article][PubMed]
    [Google Scholar]
  37. Post P. R., Carvalho R., Galler R. 1991; Glycosylation and secretion of yellow fever virus nonstructural protein NS1. Virus Res 18:291–302 [View Article][PubMed]
    [Google Scholar]
  38. Pryor M. J., Wright P. J. 1994; Glycosylation mutants of dengue virus NS1 protein. J Gen Virol 75:1183–1187 [View Article][PubMed]
    [Google Scholar]
  39. Rossi S. L., Ross T. M., Evans J. D. 2010; West Nile virus. Clin Lab Med 30:47–65 [View Article][PubMed]
    [Google Scholar]
  40. Scheres S. H., Valle M., Nuñez R., Sorzano C. O., Marabini R., Herman G. T., Carazo J. M. 2005; Maximum-likelihood multi-reference refinement for electron microscopy images. J Mol Biol 348:139–149 [View Article][PubMed]
    [Google Scholar]
  41. Schlesinger J. J., Brandriss M. W., Walsh E. E. 1987; Protection of mice against dengue 2 virus encephalitis by immunization with the dengue 2 virus non-structural glycoprotein NS1. J Gen Virol 68:853–857 [View Article][PubMed]
    [Google Scholar]
  42. Schmidt A. C. 2010; Response to dengue fever – the good, the bad, and the ugly?. N Engl J Med 363:484–487 [View Article][PubMed]
    [Google Scholar]
  43. Smith G. W., Wright P. J. 1985; Synthesis of proteins and glycoproteins in dengue type 2 virus-infected Vero and Aedes albopictus cells. J Gen Virol 66:559–571 [View Article][PubMed]
    [Google Scholar]
  44. Sorzano C. O., de la Fraga L. G., Clackdoyle R., Carazo J. M. 2004; Normalizing projection images: a study of image normalizing procedures for single particle three-dimensional electron microscopy. Ultramicroscopy 101:129–138 [View Article][PubMed]
    [Google Scholar]
  45. Sun D. S., King C. C., Huang H. S., Shih Y. L., Lee C. C., Tsai W. J., Yu C. C., Chang H. H. 2007; Antiplatelet autoantibodies elicited by dengue virus non-structural protein 1 cause thrombocytopenia and mortality in mice. J Thromb Haemost 5:2291–2299 [View Article][PubMed]
    [Google Scholar]
  46. Typke D., Nordmeyer R. A., Jones A., Lee J., Avila-Sakar A., Downing K. H., Glaeser R. M. 2005; High-throughput film-densitometry: an efficient approach to generate large data sets. J Struct Biol 149:17–29 [View Article][PubMed]
    [Google Scholar]
  47. Wallis T. P., Huang C. Y., Nimkar S. B., Young P. R., Gorman J. J. 2004; Determination of the disulfide bond arrangement of dengue virus NS1 protein. J Biol Chem 279:20729–20741 [View Article][PubMed]
    [Google Scholar]
  48. Westaway E. G., Goodman M. R. 1987; Variation in distribution of the three flavivirus-specified glycoproteins detected by immunofluorescence in infected Vero cells. Arch Virol 94:215–228 [View Article][PubMed]
    [Google Scholar]
  49. Westaway E. G., Mackenzie J. M., Kenney M. T., Jones M. K., Khromykh A. A. 1997; Ultrastructure of Kunjin virus-infected cells: colocalization of NS1 and NS3 with double-stranded RNA, and of NS2B with NS3, in virus-induced membrane structures. J Virol 71:6650–6661[PubMed]
    [Google Scholar]
  50. Winkler G., Randolph V. B., Cleaves G. R., Ryan T. E., Stollar V. 1988; Evidence that the mature form of the flavivirus nonstructural protein NS1 is a dimer. Virology 162:187–196 [View Article][PubMed]
    [Google Scholar]
  51. Woolford D., Ericksson G., Rothnagel R., Muller D., Landsberg M. J., Pantelic R. S., McDowall A., Pailthorpe B., Young P. R.other authors 2007; SwarmPS: rapid, semi-automated single particle selection software. J Struct Biol 157:174–188 [View Article][PubMed]
    [Google Scholar]
  52. Wright P. J., Cauchi M. R., Ng M. L. 1989; Definition of the carboxy termini of the three glycoproteins specified by dengue virus type 2. Virology 171:61–67 [View Article][PubMed]
    [Google Scholar]
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