Functional chimeric HN glycoproteins derived from Newcastle disease virus and human parainfluenza virus-3

  • R. Deng
  • A. M. Mirza
  • P. J. Mahon
  • R. M. Iorio


Newcastle disease virus (NDV) is primarily a respiratory tract pathogen of birds, particularly chickens, but it occasionally produces infection in man. Human parainfluenza virus type 3 (hPIV3) is a common respiratory pathogen, particularly in young children. These two viruses gain entry to host cells via direct fusion between the viral envelope and the cell membrane, mediated by the two surface glycoproteins: the hemagglutinin-neuraminidase (HN) and fusion (F) proteins. Promotion of fusion by HN and F requires that they are derived from homologous viruses. We have constructed chimeric proteins composed of domains from heterologous HN proteins. Their ability to bind cellular receptors and to complement the F protein of each virus in the promotion of fusion were evaluated in a transient expression system. The fusion specificity was found to segregate with a segment extending from the middle of the transmembrane anchor to the top of the putative stalk region of the ectodomain. All of the chimeras, in which the globular domain is derived from the NDV HN and various lengths of the stalk region are derived from the hPIV3 HN maintain receptor binding activity, but some have markedly reduced neuraminidase (NA) activity. Decrease in the NA activity of the chimeras correlates with alteration in the antigenic structure of the globular domain. This suggests that the stalk region of the HN spike is important for maintenance of the structure and function of the globular domain of the HN protein spike.


Newcastle Disease Virus Globular Domain Sendai Virus Globular Head Receptor Binding Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Alexander DJ (1988) Newcastle disease: methods of spread. In: Alexander DJ (ed) Newcastle disease. Kluwer, Norwell, pp 262–272CrossRefGoogle Scholar
  2. 2.
    Bousse T, Takimoto T, Gorman WL, Takahashi T, Portner A (1994) Regions on the hemagglutinin-neuraminidase proteins of human parainfluenza virus type-1 and Sendai virus important for membrane fusion. Virology 204: 506–514PubMedCrossRefGoogle Scholar
  3. 3.
    Cattaneo R, Rose JK (1993) Cell fusion by the envelope glycoproteins of persistent measles viruses which caused lethal human brain disease. J Virol 67: 1493–1502PubMedGoogle Scholar
  4. 4.
    Choppin PW, Scheid A (1980) The role of viral glycoproteins in adsorption penetration and pathogenicity of viruses. Rev Infect Dis 2: 40–61PubMedCrossRefGoogle Scholar
  5. 5.
    Coelingh KJ, Winter CC, Murphy BR, Rice JM, Kimball PC, Olmsted RA, Collins PL (1986) Conserved epitopes on the hemagglutinin-neuraminidase proteins of human and bovine parainfluenza type 3 viruses: nucleotide sequence analysis of variants selected with monoclonal antibodies. J Virol 60: 90–96PubMedGoogle Scholar
  6. 6.
    Coelingh KV, Winter CC, Jorgenson ED, Murphy BR (1987) Antigenic and structural properties of the hemagglutinin-neuraminidase glycoprotein of human parainfluenza type 3: sequence analysis of variants selected with monoclonal antibodies which inhibit infectivity, hemagglutination and neuraminidase activities. J Virol 61: 1473–1477Google Scholar
  7. 7.
    Colman PM, Hoyne PA, Lawrence MC (1993) Sequence and structure alignment of paramyxovirus hemagglutinin-neuraminidase with influenza virus neuraminidase. J Virol 67: 2972–2980PubMedGoogle Scholar
  8. 8.
    Compans RW, Dimmock NJ, Meier-Ewert H (1969) Effect of antibody to neuraminidase on the maturation and hemagglutinating activity of an influenza A2 virus. J Virol 4: 528–534PubMedGoogle Scholar
  9. 9.
    Deng R, Wang Z, Glickman RL, Iorio RM (1994) Glycosylation within an antigenic site on the HN glycoprotein of Newcastle disease virus interferes with its role in the promotion of membrane fusion. Virology 204: 17–26PubMedCrossRefGoogle Scholar
  10. 10.
    Deng R, Wang Z, Mirza AM, Iorio RM (1995) Localization of a domain on the paramyxovirus attachment protein required for the promotion of cellular fusion by its homologous fusion protein spike. Virology 209: 457–469PubMedCrossRefGoogle Scholar
  11. 11.
    Fuerst TR, Niles EG, Studier FW, Moss B (1986) Eucaryotic transient expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase, Proc Natl Acad Sci USA 83: 8122–8126PubMedCrossRefGoogle Scholar
  12. 12.
    Ginsberg HG (1988) Paramyxoviruses. In: Dulbecco R, Ginsberg HS (eds) Virology, 2nd ed. JB Lippincott, Philadelphia, pp 239–259Google Scholar
  13. 13.
    Heminway BH, Yu Y, Galinski MS (1994) Both surface glycoproteins are necessary for human parainfluenza type 3 mediated cell fusion. Virus Res 31: 1–16PubMedCrossRefGoogle Scholar
  14. 14.
    Horvath CM, Paterson RG, Shaughnessy MA, Wood R, Lamb RA (1992) Biological activity of paramyxovirus fusion proteins: factors influencing formation of syncytia. J Virol 66: 4564–4569PubMedGoogle Scholar
  15. 15.
    Hu X, Ray R, Compans RW (1992) Functional interactions between the fusion protein and hemagglutinin-neuraminidase of human parainfluenza viruses. J Virol 66: 1528–1534PubMedGoogle Scholar
  16. 16.
    Huberman K, Peluso RW, Moscona A (1995) Hemagglutinin-neuraminidase of human parainfluenza 3: role of the neuraminidase in the viral life cycle. Virology 214: 294–300PubMedCrossRefGoogle Scholar
  17. 17.
    Iorio RM, Bratt MA (1984) Monoclonal antibodies as functional probes of the HN glycoprotein of Newcastle disease virus: antigenic separation of the hemagglutinating and neuraminidase sites. J Immunol 133: 2215–2219PubMedGoogle Scholar
  18. 18.
    Iorio RM, Glickman RL, Riel AM, Sheehan JP, Bratt MA (1989) Functional and neutralization profile of seven overlapping antigenic sites on the HN glycoprotein of Newcastle disease virus: monoclonal antibodies to some sites prevent viral attachment. J Virus Res 13: 245–262CrossRefGoogle Scholar
  19. 19.
    Iorio RM, Syddall RJ, Sheehan JP, Bratt MA, Glickman RL, Riel AM (1991) Neutralization map of the HN glycoprotein of Newcastle disease virus: domains recognized by monoclonal antibodies that prevent receptor recognition. J Virol 65: 4999–5006PubMedGoogle Scholar
  20. 20.
    Lamb RA (1993) Paramyxovirus fusion: a hypothesis for changes. Virology 197: 1–11PubMedCrossRefGoogle Scholar
  21. 21.
    Lui C, Air GM (1993) Selection and characterization of a neuraminidase-minus mutant of influenza virus and its rescue by cloned neuraminidase gene. Virology 194: 403–407CrossRefGoogle Scholar
  22. 22.
    Lui C, Eichelberger MC, Compans RW, Air GM (1995) Influenza type A virus neuraminidase does not play a role in viral entry, replication, assembly, or budding. J Virol 69: 1099–1106Google Scholar
  23. 23.
    Mahon PJ, Deng R, Mirza AM, Iorio RM (1995) Cooperative neuraminidase activity in a paramyxovirus. Virology 213: 241–244PubMedCrossRefGoogle Scholar
  24. 24.
    Malvoisin E, Wild TF (1993) Measles virus glycoproteins: studies on the structure and interaction of the haemagglutinin and fusion proteins. J Gen Virol 74: 2365–2372PubMedCrossRefGoogle Scholar
  25. 25.
    Mirza AM, Sheehan JP, Hardy LW, Glickman RL, Iorio RM (1993) Structure and function of a membrane anchor-less form of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus. J Biol Chem 268: 21425–21431PubMedGoogle Scholar
  26. 26.
    Mirza AM, Deng R, Iorio RM (1994) Site-directed mutagenesis of a conserved hexa-peptide in the paramyxovirus hemagglutinin-neuraminidase glycoprotein: effects on antigenic structure and function. J Virol 68: 5093–5099PubMedGoogle Scholar
  27. 27.
    Morrison T, Portner A (1991) Structure, function, and intracellular processing of the glycoproteins of paramyxoviridae. In: Kingsbury DW (ed) The paramyxoviruses. Plenum, New York, pp 347–382CrossRefGoogle Scholar
  28. 28.
    Moscona A, Peluso RW (1991) Fusion properties of cells infected with human parainfluenza virus type 3: receptor requirements for viral spread and virus-mediated membrane fusion. J Virol 66: 6280–6287Google Scholar
  29. 29.
    Palese P, Compans RW (1976) Inhibition of influenza virus replication in tissue culture by 2-deoxy-2,3-dehydro-N-trifluoroacetylneuraminic acid (FANA): mechanism of action. J Gen Virol 33: 159–163PubMedCrossRefGoogle Scholar
  30. 30.
    Portner A (1981) Evidence for two different sites in the HN glycoprotein involved in neuraminidase and hemagglutinating activities. In: Bishop DHL, Compans RW (eds) The replication of negative-strand viruses. Elsevier/North-Holland, New York, pp 465–470Google Scholar
  31. 31.
    Portner A, Scroggs RA, Metzger DW (1987) Distinct functions of antigenic sites of the HN glycoprotein of Sendai virus. Virology 158: 61–68PubMedCrossRefGoogle Scholar
  32. 32.
    Ray R, Compans RW (1986) Monoclonal antibodies reveal extensive antigenic differences between the hemagglutinin-neuraminidase glycoproteins of human and bovine parainfluenza 3 viruses. Virology 148: 232–236PubMedCrossRefGoogle Scholar
  33. 33.
    Scheid A, Choppin PW (1974) Identification and biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis and infectivity by proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology 57: 375–490CrossRefGoogle Scholar
  34. 34.
    Sergei T, McGinnes LW, Peeples MA, Morrison TG (1993) The attachment function of the Newcastle disease virus hemagglutinin-neuraminidase protein can be separated from fusion promotion by mutation. Virology 193: 717–726CrossRefGoogle Scholar
  35. 35.
    Sheehan JP, Iorio RM (1992) A single amino acid substitution in the hemagglutinin-neuraminidase of Newcastle disease virus results in a protein deficient in both functions. Virology 189: 778–781PubMedCrossRefGoogle Scholar
  36. 36.
    Schuy W, Garten W, Linder D, Klenk HD (1984) The carboxyterminus of the hemagglutinin-neuraminidase of Newcastle disease virus is exposed at the surface of the viral envelope. Virus Res 1: 415–426PubMedCrossRefGoogle Scholar
  37. 37.
    Smith GW, Hightower LE (1980) Uncoupling of the hemagglutinating and neuraminidase activities of Newcastle disease virus (NDV). In: Fields B, Jaenish R, Fox CF (eds) Animal virus genetics. Academic Press, New York, pp 623–632Google Scholar
  38. 38.
    Smith GW, Hightower LE (1982) Revertant analysis of a temperature-sensitive mutant of Newcastle disease virus with defective glycoprotein: Implications of the fusion glycoprotein in cell killing and isolation of a neuraminidase-deficient hemagglutinating virus. J Virol 42: 659–668PubMedGoogle Scholar
  39. 39.
    Tababayashi K, Compans RW (1996) Functional interaction of paramyxovirus glycoproteins: identification of a domain in Sendai virus HN which promotes cell fusion. J Virol 70: 6112–6118Google Scholar
  40. 40.
    Tanaka Y, Heminway BR, Galinski MS (1996) Down-regulation of paramyxovirus hemagglutinin-neuraminidase glycoprotein surface expression by a mutant fusion protein containing a retention signal for the endoplasmic reticulum. J Virol 71: 5005–5015Google Scholar
  41. 41.
    Thompson SD, Laver WG, Murti KG, Portner A (1988) Isolation of a biologically active soluble form of the hemagglutinin-neuraminidase protein of Sendai virus. J Virol 62:4653–4660PubMedGoogle Scholar
  42. 42.
    Tsurodome 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: 190–203CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 1997

Authors and Affiliations

  • R. Deng
    • 1
    • 3
  • A. M. Mirza
    • 1
  • P. J. Mahon
    • 2
  • R. M. Iorio
    • 1
  1. 1.Department of Molecular Genetics and MicrobiologyUniversity of Massachusetts Medical SchoolWorcesterUSA
  2. 2.Department of BiologyAssumption CollegeWorcesterUSA
  3. 3.Molecular and Cellular Virology, Animal Health Biological Discovery, Central Research DivisionPfizer Inc.GrotonUSA

Personalised recommendations