1887

Abstract

Since the discovery of a non-virulent infectious salmon anaemia virus (ISAV) HPR0 variant, many studies have speculated on the functional role of deletions within the highly polymorphic region (HPR) of genomic segment 6, which codes for the haemagglutinin–esterase (HE) protein. To address this issue, mutant HE proteins with deletions in their HPR were generated from the Scottish HPR0 template (NWM10) and fusion-inducing activity was measured using lipid (octadecyl rhodamine B) and content mixing assays (firefly luciferase). Segment six HPR was found to have a strong influence on ISAV fusion, and deletions in this near-membrane region predominantly increased the fusion-inducing ability of the resulting HE proteins. The position and length of the HPR deletions were not significant factors, suggesting that they may affect fusion non-specifically. In comparison, the amino acid composition of the associated fusion (F) protein was a more crucial criterion. Antibody co-patching and confocal fluorescence demonstrated that the HE and F proteins were highly co-localized, forming defined clusters on the cell surface post-transfection. The binding of erythrocyte ghosts on the attachment protein caused a reduction in the percentage of co-localization, suggesting that ISAV fusion might be triggered through physical separation of the F and HE proteins. In this process, HPR deletion appeared to modulate and reduce the strength of interaction between the two glycoproteins, causing more F protein to be released and activated. This work provides a first insight into the mechanism of virulence acquisition through HPR deletion, with fusion enhancement acting as a major contributing factor.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.061648-0
2014-05-01
2019-12-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/95/5/1015.html?itemId=/content/journal/jgv/10.1099/vir.0.061648-0&mimeType=html&fmt=ahah

References

  1. Aguilar H. C. , Matreyek K. A. , Filone C. M. , Hashimi S. T. , Levroney E. L. , Negrete O. A. , Bertolotti-Ciarlet A. , Choi D. Y. , McHardy I. . & other authors ( 2006; ). N-glycans on Nipah virus fusion protein protect against neutralization but reduce membrane fusion and viral entry. . J Virol 80:, 4878–4889. [CrossRef] [PubMed]
    [Google Scholar]
  2. Aguilar H. C. , Matreyek K. A. , Choi D. Y. , Filone C. M. , Young S. , Lee B. . ( 2007; ). Polybasic KKR motif in the cytoplasmic tail of Nipah virus fusion protein modulates membrane fusion by inside-out signaling. . J Virol 81:, 4520–4532. [CrossRef] [PubMed]
    [Google Scholar]
  3. Armstrong R. T. , Kushnir A. S. , White J. M. . ( 2000; ). The transmembrane domain of influenza hemagglutinin exhibits a stringent length requirement to support the hemifusion to fusion transition. . J Cell Biol 151:, 425–437. [CrossRef] [PubMed]
    [Google Scholar]
  4. Aspehaug V. , Mikalsen A. B. , Snow M. , Biering E. , Villoing S. . ( 2005; ). Characterization of the infectious salmon anemia virus fusion protein. . J Virol 79:, 12544–12553. [CrossRef] [PubMed]
    [Google Scholar]
  5. Ayllón J. , Villar E. , Muñoz-Barroso I. . ( 2010; ). Mutations in the ectodomain of newcastle disease virus fusion protein confer a hemagglutinin–neuraminidase-independent phenotype. . J Virol 84:, 1066–1075. [CrossRef] [PubMed]
    [Google Scholar]
  6. Berggård T. , Linse S. , James P. . ( 2007; ). Methods for the detection and analysis of protein–protein interactions. . Proteomics 7:, 2833–2842. [CrossRef] [PubMed]
    [Google Scholar]
  7. Bouchard D. , Keleher W. , Opitz H. M. , Blake S. , Edwards K. C. , Nicholson B. L. . ( 1999; ). Isolation of infectious salmon anemia virus (ISAV) from Atlantic salmon in New Brunswick, Canada. . Dis Aquat Organ 35:, 131–137. [CrossRef] [PubMed]
    [Google Scholar]
  8. Brindley M. A. , Plemper R. K. . ( 2010; ). Blue native PAGE and biomolecular complementation reveal a tetrameric or higher-order oligomer organization of the physiological measles virus attachment protein H. . J Virol 84:, 12174–12184. [CrossRef] [PubMed]
    [Google Scholar]
  9. Brindley M. A. , Takeda M. , Plattet P. , Plemper R. K. . ( 2012; ). Triggering the measles virus membrane fusion machinery. . Proc Natl Acad Sci U S A 109:, E3018–E3027. [CrossRef] [PubMed]
    [Google Scholar]
  10. Byun H. M. , Suh D. , Yoon H. , Kim J. M. , Choi H. G. , Kim W. K. , Ko J. J. , Oh Y. K. . ( 2004; ). Erythrocyte ghost-mediated gene delivery for prolonged and blood-targeted expression. . Gene Ther 11:, 492–496. [CrossRef] [PubMed]
    [Google Scholar]
  11. Chang D. K. , Cheng S. F. , Kantchev E. A. , Lin C. H. , Liu Y. T. . ( 2008; ). Membrane interaction and structure of the transmembrane domain of influenza hemagglutinin and its fusion peptide complex. . BMC Biol 6:, 2. [CrossRef] [PubMed]
    [Google Scholar]
  12. Christiansen D. H. , Østergaard P. S. , Snow M. , Dale O. B. , Falk K. . ( 2011; ). A low-pathogenic variant of infectious salmon anemia virus (ISAV-HPR0) is highly prevalent and causes a non-clinical transient infection in farmed Atlantic salmon (Salmo salar L.) in the Faroe Islands. . J Gen Virol 92:, 909–918. [CrossRef] [PubMed]
    [Google Scholar]
  13. Collet B. , Secombes C. J. . ( 2005; ). Construction and analysis of a secreting expression vector for fish cells. . Vaccine 23:, 1534–1539. [CrossRef] [PubMed]
    [Google Scholar]
  14. Cook-Versloot M. , Griffiths S. , Cusack R. , McGeachy S. , Ritchie R. . ( 2004; ). Identification and characterisation of infectious salmon anaemia virus (ISAV) haemagglutinin gene highly polymorphic region (HPR) type 0 in North America. . Bull Eur Assoc Fish Pathol 24:, 203–208.
    [Google Scholar]
  15. Corey E. A. , Iorio R. M. . ( 2007; ). Mutations in the stalk of the measles virus hemagglutinin protein decrease fusion but do not interfere with virus-specific interaction with the homologous fusion protein. . J Virol 81:, 9900–9910. [CrossRef] [PubMed]
    [Google Scholar]
  16. Cunningham C. O. , Gregory A. , Black J. , Simpson I. , Raynard R. S. . ( 2002; ). A novel variant of the infectious salmon anaemia virus (ISAV) haemagglutinin gene suggests mechanisms for virus diversity. . Bull Eur Assoc Fish Pathol 22:, 366–374.
    [Google Scholar]
  17. Devold M. , Falk K. , Dale B. , Krossøy B. , Biering E. , Aspehaug V. , Nilsen F. , Nylund A. . ( 2001; ). Strain variation, based on the hemagglutinin gene, in Norwegian ISA virus isolates collected from 1987 to 2001: indications of recombination. . Dis Aquat Organ 47:, 119–128. [CrossRef] [PubMed]
    [Google Scholar]
  18. Falk K. , Namork E. , Rimstad E. , Mjaaland S. , Dannevig B. H. . ( 1997; ). Characterization of infectious salmon anemia virus, an orthomyxo-like virus isolated from Atlantic salmon (Salmo salar L.). . J Virol 71:, 9016–9023.[PubMed]
    [Google Scholar]
  19. Falk K. , Namork E. , Dannevig B. H. . ( 1998; ). Characterization and applications of a monoclonal antibody against infectious salmon anaemia virus. . Dis Aquat Organ 34:, 77–85. [CrossRef] [PubMed]
    [Google Scholar]
  20. Falk K. , Aspehaug V. , Vlasak R. , Endresen C. . ( 2004; ). Identification and characterization of viral structural proteins of infectious salmon anemia virus. . J Virol 78:, 3063–3071. [CrossRef] [PubMed]
    [Google Scholar]
  21. Gilboa L. , Nohe A. , Geissendörfer T. , Sebald W. , Henis Y. I. , Knaus P. . ( 2000; ). Bone morphogenetic protein receptor complexes on the surface of live cells: a new oligomerization mode for serine/threonine kinase receptors. . Mol Biol Cell 11:, 1023–1035. [CrossRef] [PubMed]
    [Google Scholar]
  22. Godoy M. G. , Aedo A. , Kibenge M. J. , Groman D. B. , Yason C. V. , Grothusen H. , Lisperguer A. , Calbucura M. , Avendaño F. . & other authors ( 2008; ). First detection, isolation and molecular characterization of infectious salmon anaemia virus associated with clinical disease in farmed Atlantic salmon (Salmo salar) in Chile. . BMC Vet Res 4:, 28. [CrossRef] [PubMed]
    [Google Scholar]
  23. Gravel K. A. , Morrison T. G. . ( 2003; ). Interacting domains of the HN and F proteins of Newcastle disease virus. . J Virol 77:, 11040–11049. [CrossRef] [PubMed]
    [Google Scholar]
  24. Hellebø A. , Vilas U. , Falk K. , Vlasak R. . ( 2004; ). Infectious salmon anemia virus specifically binds to and hydrolyzes 4-O-acetylated sialic acids. . J Virol 78:, 3055–3062. [CrossRef] [PubMed]
    [Google Scholar]
  25. Hoekstra D. , de Boer T. , Klappe K. , Wilschut J. . ( 1984; ). Fluorescence method for measuring the kinetics of fusion between biological membranes. . Biochemistry 23:, 5675–5681. [CrossRef] [PubMed]
    [Google Scholar]
  26. Hotchin J. E. , Cohen S. M. , Ruska H. , Ruska C. . ( 1958; ). Electron microscopical aspects of hemadsorption in tissue cultures infected with influenza virus. . Virology 6:, 689–701. [CrossRef] [PubMed]
    [Google Scholar]
  27. Johnson A. , Binette S. , Coo-Versloot M. , Beattie M. , McGeachy S. , Gagne N. , McDonald J. T. , Ritchie R. . ( 2008; ). Association between ISAV Mortalities and ISAV Molecular Type in the Bay of Fundy, Canada. . Can Tech Report Fish Aquat Sci 2782. Canada:.
    [Google Scholar]
  28. Kemble G. W. , Danieli T. , White J. M. . ( 1994; ). Lipid-anchored influenza hemagglutinin promotes hemifusion, not complete fusion. . Cell 76:, 383–391. [CrossRef] [PubMed]
    [Google Scholar]
  29. Keren T. , Roth M. G. , Henis Y. I. . ( 2001; ). Internalization-competent influenza hemagglutinin mutants form complexes with clathrin-deficient multivalent AP-2 oligomers in live cells. . J Biol Chem 276:, 28356–28363. [CrossRef] [PubMed]
    [Google Scholar]
  30. Kibenge F. S. , Gárate O. N. , Johnson G. , Arriagada R. , Kibenge M. J. , Wadowska D. . ( 2001; ). Isolation and identification of infectious salmon anaemia virus (ISAV) from Coho salmon in Chile. . Dis Aquat Organ 45:, 9–18. [CrossRef] [PubMed]
    [Google Scholar]
  31. Kibenge F. S. , Munir K. , Kibenge M. J. , Joseph T. , Moneke E. . ( 2004; ). Infectious salmon anemia virus: causative agent, pathogenesis and immunity. . Anim Health Res Rev 5:, 65–78. [CrossRef] [PubMed]
    [Google Scholar]
  32. Kibenge F. S. , Kibenge M. J. , Wang Y. , Qian B. , Hariharan S. , McGeachy S. . ( 2007; ). Mapping of putative virulence motifs on infectious salmon anemia virus surface glycoprotein genes. . J Gen Virol 88:, 3100–3111. [CrossRef] [PubMed]
    [Google Scholar]
  33. Kibenge F. S. , Godoy M. G. , Wang Y. , Kibenge M. J. , Gherardelli V. , Mansilla S. , Lisperger A. , Jarpa M. , Larroquete G. . & other authors ( 2009; ). Infectious salmon anaemia virus (ISAV) isolated from the ISA disease outbreaks in Chile diverged from ISAV isolates from Norway around 1996 and was disseminated around 2005, based on surface glycoprotein gene sequences. . Virol J 6:, 88. [CrossRef] [PubMed]
    [Google Scholar]
  34. Krossøy B. , Devold M. , Sanders L. , Knappskog P. M. , Aspehaug V. , Falk K. , Nylund A. , Koumans S. , Endresen C. , Biering E. . ( 2001; ). Cloning and identification of the infectious salmon anaemia virus haemagglutinin. . J Gen Virol 82:, 1757–1765.[PubMed]
    [Google Scholar]
  35. Lamb R. A. , Kolakofsky D. . ( 2001; ). Paramyxoviridae: the viruses and their replication. . In Fundamental Virology, pp. 1305–1340. Edited by Fields B. N. , Knipe D. M. , Kato A. . . New York:: Lippencott-Raven;.
    [Google Scholar]
  36. Lee J. K. , Prussia A. , Paal T. , White L. K. , Snyder J. P. , Plemper R. K. . ( 2008; ). Functional interaction between paramyxovirus fusion and attachment proteins. . J Biol Chem 283:, 16561–16572. [CrossRef] [PubMed]
    [Google Scholar]
  37. Li J. , Quinlan E. , Mirza A. , Iorio R. M. . ( 2004; ). Mutated form of the Newcastle disease virus hemagglutinin–neuraminidase interacts with the homologous fusion protein despite deficiencies in both receptor recognition and fusion promotion. . J Virol 78:, 5299–5310. [CrossRef] [PubMed]
    [Google Scholar]
  38. Markosyan R. M. , Cohen F. S. , Melikyan G. B. . ( 2000; ). The lipid-anchored ectodomain of influenza virus hemagglutinin (GPI-HA) is capable of inducing nonenlarging fusion pores. . Mol Biol Cell 11:, 1143–1152. [CrossRef] [PubMed]
    [Google Scholar]
  39. McBeath A. J. , Bain N. , Snow M. . ( 2009; ). Surveillance for infectious salmon anaemia virus HPR0 in marine Atlantic salmon farms across Scotland. . Dis Aquat Organ 87:, 161–169. [CrossRef] [PubMed]
    [Google Scholar]
  40. McBeath A. , Fourrier M. , Munro E. , Falk K. , Snow M. . ( 2011; ). Presence of a full-length highly polymorphic region (HPR) in the ISAV haemagglutinin-esterase does not affect the primary functions of receptor binding and esterase activity. . Arch Virol 156:, 2285–2289. [CrossRef] [PubMed]
    [Google Scholar]
  41. Melanson V. R. , Iorio R. M. . ( 2004; ). Amino acid substitutions in the F-specific domain in the stalk of the Newcastle disease virus HN protein modulate fusion and interfere with its interaction with the F protein. . J Virol 78:, 13053–13061. [CrossRef] [PubMed]
    [Google Scholar]
  42. Melikyan G. B. , Lin S. , Roth M. G. , Cohen F. S. . ( 1999; ). Amino acid sequence requirements of the transmembrane and cytoplasmic domains of influenza virus hemagglutinin for viable membrane fusion. . Mol Biol Cell 10:, 1821–1836. [CrossRef] [PubMed]
    [Google Scholar]
  43. Mikalsen A. B. , Sindre H. , Mjaaland S. , Rimstad E. . ( 2005; ). Expression, antigenicity and studies on cell receptor binding of the hemagglutinin of infectious salmon anemia virus. . Arch Virol 150:, 1621–1637. [CrossRef] [PubMed]
    [Google Scholar]
  44. Mjaaland S. , Rimstad E. , Falk K. , Dannevig B. H. . ( 1997; ). Genomic characterization of the virus causing infectious salmon anemia in Atlantic salmon (Salmo salar L.): an orthomyxo-like virus in a teleost. . J Virol 71:, 7681–7686.[PubMed]
    [Google Scholar]
  45. Mjaaland S. , Hungnes O. , Teig A. , Dannevig B. H. , Thorud K. , Rimstad E. . ( 2002; ). Polymorphism in the infectious salmon anemia virus hemagglutinin gene: importance and possible implications for evolution and ecology of infectious salmon anemia disease. . Virology 304:, 379–391. [CrossRef] [PubMed]
    [Google Scholar]
  46. Mullins J. E. , Groman D. B. , Wadowska D. . ( 1998; ). Infectious salmon anaemia in salt water Atlantic salmon (Salmo salar L.) in New Brunswick, Canada. . Bull Eur Assoc Fish Pathol 18:, 110–114.
    [Google Scholar]
  47. Nylund A. , Devold M. , Plarre H. , Isdal E. , Aarseth M. . ( 2003; ). Emergence and maintenance of infectious salmon anaemia virus (ISAV) in Europe: a new hypothesis. . Dis Aquat Organ 56:, 11–24. [CrossRef] [PubMed]
    [Google Scholar]
  48. Nylund A. , Plarre H. , Karlsen M. , Fridell F. , Ottem K. F. , Bratland A. , Saether P. A. . ( 2007; ). Transmission of infectious salmon anaemia virus (ISAV) in farmed populations of Atlantic salmon (Salmo salar). . Arch Virol 152:, 151–179. [CrossRef] [PubMed]
    [Google Scholar]
  49. Ohuchi M. , Asaoka N. , Sakai T. , Ohuchi R. . ( 2006; ). Roles of neuraminidase in the initial stage of influenza virus infection. . Microbes Infect 8:, 1287–1293. [CrossRef] [PubMed]
    [Google Scholar]
  50. Okada C. Y. , Rechsteiner M. . ( 1982; ). Introduction of macromolecules into cultured mammalian cells by osmotic lysis of pinocytic vesicles. . Cell 29:, 33–41. [CrossRef] [PubMed]
    [Google Scholar]
  51. Paal T. , Brindley M. A. , St Clair C. , Prussia A. , Gaus D. , Krumm S. A. , Snyder J. P. , Plemper R. K. . ( 2009; ). Probing the spatial organization of measles virus fusion complexes. . J Virol 83:, 10480–10493. [CrossRef] [PubMed]
    [Google Scholar]
  52. Palese P. , Shaw M. L. . ( 2007; ). Orthomyxoviridae: the viruses and their replication. . In Fields Virology, pp. 1647–1690. Edited by Knipe D. M. , Howley P. M. . . Philadelphia:: Lippincott, Williams & Wilkins;.
    [Google Scholar]
  53. Plemper R. K. , Hammond A. L. , Cattaneo R. . ( 2001; ). Measles virus envelope glycoproteins hetero-oligomerize in the endoplasmic reticulum. . J Biol Chem 276:, 44239–44246. [CrossRef] [PubMed]
    [Google Scholar]
  54. Plemper R. K. , Hammond A. L. , Gerlier D. , Fielding A. K. , Cattaneo R. . ( 2002; ). Strength of envelope protein interaction modulates cytopathicity of measles virus. . J Virol 76:, 5051–5061. [CrossRef] [PubMed]
    [Google Scholar]
  55. Plemper R. K. , Brindley M. A. , Iorio R. M. . ( 2011; ). Structural and mechanistic studies of measles virus illuminate paramyxovirus entry. . PLoS Pathog 7:, e1002058. [CrossRef] [PubMed]
    [Google Scholar]
  56. Rimstad E. , Mjaaland S. , Snow M. , Mikalsen A. B. , Cunningham C. O. . ( 2001; ). Characterization of the infectious salmon anemia virus genomic segment that encodes the putative hemagglutinin. . J Virol 75:, 5352–5356. [CrossRef] [PubMed]
    [Google Scholar]
  57. Scyth B. D. , Olesen N. J. , Østergård P. , Falk K. . ( 2003; ). Laboratory diagnosis of infectious salmon anaemia (ISA): experience gained from the outbreaks on the Faroe Islands 2000–2003. . In 11th International Conference of the EAFP – Diseases of Fish and Shellfish, 21–26 September, Malta.
    [Google Scholar]
  58. Shvartsman D. E. , Kotler M. , Tall R. D. , Roth M. G. , Henis Y. I. . ( 2003; ). Differently anchored influenza hemagglutinin mutants display distinct interaction dynamics with mutual rafts. . J Cell Biol 163:, 879–888. [CrossRef] [PubMed]
    [Google Scholar]
  59. Tanabayashi K. , Compans R. W. . ( 1996; ). Functional interaction of paramyxovirus glycoproteins: identification of a domain in Sendai virus HN which promotes cell fusion. . J Virol 70:, 6112–6118.[PubMed]
    [Google Scholar]
  60. Thorud K. , Djupvik H. O. . ( 1988; ). Infectious anaemia in Atlantic salmon (Salmo salar L.). . Bull Eur Assoc Fish Pathol 8:, 109–111.
    [Google Scholar]
  61. Tsurudome M. , Kawano M. , Yuasa T. , Tabata N. , Nishio M. , Komada H. , Ito Y. . ( 1995; ). Identification of regions on the hemagglutinin–neuraminidase protein of human parainfluenza virus type 2 important for promoting cell fusion. . Virology 213:, 190–203. [CrossRef] [PubMed]
    [Google Scholar]
  62. Wang X. , Zhang G. , Zhao J. , Wang M. . ( 2005; ). Interacting domains of the HN and F protein of paramyxovirus. . Chin Sci Bull 50:, 2598–2601.[CrossRef]
    [Google Scholar]
  63. White J. M. , Delos S. E. , Brecher M. , Schornberg K. . ( 2008; ). Structures and mechanisms of viral membrane fusion proteins: multiple variations on a common theme. . Crit Rev Biochem Mol Biol 43:, 189–219. [CrossRef] [PubMed]
    [Google Scholar]
  64. Zhou J. , Dutch R. E. , Lamb R. A. . ( 1997; ). Proper spacing between heptad repeat B and the transmembrane domain boundary of the paramyxovirus SV5 F protein is critical for biological activity. . Virology 239:, 327–339. [CrossRef] [PubMed]
    [Google Scholar]
  65. Zimmer G. , Bossow S. , Kolesnikova L. , Hinz M. , Neubert W. J. , Herrler G. . ( 2005; ). A chimeric respiratory syncytial virus fusion protein functionally replaces the F and HN glycoproteins in recombinant Sendai virus. . J Virol 79:, 10467–10477. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.061648-0
Loading
/content/journal/jgv/10.1099/vir.0.061648-0
Loading

Data & Media loading...

Supplements

Supplementary material 

PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error