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. 1997 Jan;71(1):650–656. doi: 10.1128/jvi.71.1.650-656.1997

Association of the parainfluenza virus fusion and hemagglutinin-neuraminidase glycoproteins on cell surfaces.

Q Yao 1, X Hu 1, R W Compans 1
PMCID: PMC191097  PMID: 8985396

Abstract

We previously observed that cell fusion caused by human parainfluenza virus type 2 or type 3 requires the expression of both the fusion (F) and hemagglutinin-neuraminidase (HN) glycoproteins from the same virus type, indicating that a type-specific interaction between F and HN is needed for the induction of cell fusion. In the present study we have further investigated the fusion properties of F and HN proteins of parainfluenza virus type 1 (PI1), type 2 (PI2), and type 3 (PI3), Sendai virus (SN), and simian virus 5 (SV5) by expression of their glycoprotein genes in HeLa T4 cells using the vaccinia virus-T7 transient expression system. Consistent with previous results, cell fusion was observed in cells transfected with homotypic F/HN proteins; with one exception, coexpression of any combination of F and HN proteins from different viruses did not result in cell fusion. The only exception was found with the closely related PI1 HN and SN HN glycoproteins, either of which could interact with SN F to induce cell fusion upon coexpression as previously reported. By specific labeling and coprecipitation of proteins expressed on the cell surface, we observed that anti-PI2 HN antiserum coprecipitated PI2 F when the homotypic PI2 F and PI2 HN were coexpressed, but not the F proteins of other paramyxoviruses when heterotypic F genes were coexpressed with PI2 HN, suggesting that the homotypic F and HN proteins are physically associated with each other on cell surfaces. Furthermore, we observed that PI3 F was found to cocap with PI3 HN but not with PI2 HN, also indicating a specific association between the homotypic proteins. These results indicate that the homotypic F and HN glycoproteins are physically associated with each other on the cell surface and suggest that such association is crucial to cell fusion induced by paramyxoviruses.

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Selected References

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  1. Bagai S., Lamb R. A. Quantitative measurement of paramyxovirus fusion: differences in requirements of glycoproteins between simian virus 5 and human parainfluenza virus 3 or Newcastle disease virus. J Virol. 1995 Nov;69(11):6712–6719. doi: 10.1128/jvi.69.11.6712-6719.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bousse T., Takimoto T., Gorman W. L., Takahashi T., Portner A. Regions on the hemagglutinin-neuraminidase proteins of human parainfluenza virus type-1 and Sendai virus important for membrane fusion. Virology. 1994 Nov 1;204(2):506–514. doi: 10.1006/viro.1994.1564. [DOI] [PubMed] [Google Scholar]
  3. Buckland R., Malvoisin E., Beauverger P., Wild F. A leucine zipper structure present in the measles virus fusion protein is not required for its tetramerization but is essential for fusion. J Gen Virol. 1992 Jul;73(Pt 7):1703–1707. doi: 10.1099/0022-1317-73-7-1703. [DOI] [PubMed] [Google Scholar]
  4. Bullough P. A., Hughson F. M., Skehel J. J., Wiley D. C. Structure of influenza haemagglutinin at the pH of membrane fusion. Nature. 1994 Sep 1;371(6492):37–43. doi: 10.1038/371037a0. [DOI] [PubMed] [Google Scholar]
  5. Carr C. M., Kim P. S. A spring-loaded mechanism for the conformational change of influenza hemagglutinin. Cell. 1993 May 21;73(4):823–832. doi: 10.1016/0092-8674(93)90260-w. [DOI] [PubMed] [Google Scholar]
  6. Cattaneo R., Rose J. K. Cell fusion by the envelope glycoproteins of persistent measles viruses which caused lethal human brain disease. J Virol. 1993 Mar;67(3):1493–1502. doi: 10.1128/jvi.67.3.1493-1502.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Choppin P. W., Scheid A. The role of viral glycoproteins in adsorption, penetration, and pathogenicity of viruses. Rev Infect Dis. 1980 Jan-Feb;2(1):40–61. doi: 10.1093/clinids/2.1.40. [DOI] [PubMed] [Google Scholar]
  8. Deng R., Wang Z., Mirza A. M., Iorio R. M. Localization of a domain on the paramyxovirus attachment protein required for the promotion of cellular fusion by its homologous fusion protein spike. Virology. 1995 Jun 1;209(2):457–469. doi: 10.1006/viro.1995.1278. [DOI] [PubMed] [Google Scholar]
  9. Ebata S. N., Côté M. J., Kang C. Y., Dimock K. The fusion and hemagglutinin-neuraminidase glycoproteins of human parainfluenza virus 3 are both required for fusion. Virology. 1991 Jul;183(1):437–441. doi: 10.1016/0042-6822(91)90162-5. [DOI] [PubMed] [Google Scholar]
  10. Fuerst T. R., Niles E. G., Studier F. W., Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122–8126. doi: 10.1073/pnas.83.21.8122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gorman W. L., Gill D. S., Scroggs R. A., Portner A. The hemagglutinin-neuraminidase glycoproteins of human parainfluenza virus type 1 and Sendai virus have high structure-function similarity with limited antigenic cross-reactivity. Virology. 1990 Mar;175(1):211–221. doi: 10.1016/0042-6822(90)90201-2. [DOI] [PubMed] [Google Scholar]
  12. Heminway B. R., Yu Y., Galinski M. S. Paramyxovirus mediated cell fusion requires co-expression of both the fusion and hemagglutinin-neuraminidase glycoproteins. Virus Res. 1994 Jan;31(1):1–16. doi: 10.1016/0168-1702(94)90066-3. [DOI] [PubMed] [Google Scholar]
  13. Homma M., Ouchi M. Trypsin action on the growth of Sendai virus in tissue culture cells. 3. Structural difference of Sendai viruses grown in eggs and tissue culture cells. J Virol. 1973 Dec;12(6):1457–1465. doi: 10.1128/jvi.12.6.1457-1465.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horvath C. M., Paterson R. G., Shaughnessy M. A., Wood R., Lamb R. A. Biological activity of paramyxovirus fusion proteins: factors influencing formation of syncytia. J Virol. 1992 Jul;66(7):4564–4569. doi: 10.1128/jvi.66.7.4564-4569.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hsu M., Scheid A., Choppin P. W. Activation of the Sendai virus fusion protein (f) involves a conformational change with exposure of a new hydrophobic region. J Biol Chem. 1981 Apr 10;256(7):3557–3563. [PubMed] [Google Scholar]
  16. Hu X. L., Ray R., Compans R. W. Functional interactions between the fusion protein and hemagglutinin-neuraminidase of human parainfluenza viruses. J Virol. 1992 Mar;66(3):1528–1534. doi: 10.1128/jvi.66.3.1528-1534.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Joseph B. S., Oldstone M. B. Antibody-induced redistribution of measles virus antigens on the cell surface. J Immunol. 1974 Oct;113(4):1205–1209. [PubMed] [Google Scholar]
  18. Kohama T., Garten W., Klenk H. D. Changes in conformation and charge paralleling proteolytic activation of Newcastle disease virus glycoproteins. Virology. 1981 Jun;111(2):364–376. doi: 10.1016/0042-6822(81)90340-8. [DOI] [PubMed] [Google Scholar]
  19. Lamb R. A. Paramyxovirus fusion: a hypothesis for changes. Virology. 1993 Nov;197(1):1–11. doi: 10.1006/viro.1993.1561. [DOI] [PubMed] [Google Scholar]
  20. Lambert D. M., Barney S., Lambert A. L., Guthrie K., Medinas R., Davis D. E., Bucy T., Erickson J., Merutka G., Petteway S. R., Jr Peptides from conserved regions of paramyxovirus fusion (F) proteins are potent inhibitors of viral fusion. Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):2186–2191. doi: 10.1073/pnas.93.5.2186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lisanti M. P., Le Bivic A., Sargiacomo M., Rodriguez-Boulan E. Steady-state distribution and biogenesis of endogenous Madin-Darby canine kidney glycoproteins: evidence for intracellular sorting and polarized cell surface delivery. J Cell Biol. 1989 Nov;109(5):2117–2127. doi: 10.1083/jcb.109.5.2117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lydy S. L., Basak S., Compans R. W. Host cell-dependent lateral mobility of viral glycoproteins. Microb Pathog. 1990 Dec;9(6):375–386. doi: 10.1016/0882-4010(90)90056-v. [DOI] [PubMed] [Google Scholar]
  23. Malvoisin E., Wild T. F. Measles virus glycoproteins: studies on the structure and interaction of the haemagglutinin and fusion proteins. J Gen Virol. 1993 Nov;74(Pt 11):2365–2372. doi: 10.1099/0022-1317-74-11-2365. [DOI] [PubMed] [Google Scholar]
  24. Marsh M., Helenius A. Virus entry into animal cells. Adv Virus Res. 1989;36:107–151. doi: 10.1016/S0065-3527(08)60583-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Matsuoka Y., Ray R., Compans R. W. Sequence of the hemagglutinin-neuraminidase gene of human parainfluenza virus type 1. Virus Res. 1990 Apr;16(1):107–113. doi: 10.1016/0168-1702(90)90047-f. [DOI] [PubMed] [Google Scholar]
  26. Morrison T., McQuain C., McGinnes L. Complementation between avirulent Newcastle disease virus and a fusion protein gene expressed from a retrovirus vector: requirements for membrane fusion. J Virol. 1991 Feb;65(2):813–822. doi: 10.1128/jvi.65.2.813-822.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moscona A., Peluso R. W. Fusion properties of cells infected with human parainfluenza virus type 3: receptor requirements for viral spread and virus-mediated membrane fusion. J Virol. 1992 Nov;66(11):6280–6287. doi: 10.1128/jvi.66.11.6280-6287.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moscona A., Peluso R. W. Fusion properties of cells persistently infected with human parainfluenza virus type 3: participation of hemagglutinin-neuraminidase in membrane fusion. J Virol. 1991 Jun;65(6):2773–2777. doi: 10.1128/jvi.65.6.2773-2777.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Paterson R. G., Shaughnessy M. A., Lamb R. A. Analysis of the relationship between cleavability of a paramyxovirus fusion protein and length of the connecting peptide. J Virol. 1989 Mar;63(3):1293–1301. doi: 10.1128/jvi.63.3.1293-1301.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rapaport D., Ovadia M., Shai Y. A synthetic peptide corresponding to a conserved heptad repeat domain is a potent inhibitor of Sendai virus-cell fusion: an emerging similarity with functional domains of other viruses. EMBO J. 1995 Nov 15;14(22):5524–5531. doi: 10.1002/j.1460-2075.1995.tb00239.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ray R., Brown V. E., Compans R. W. Glycoproteins of human parainfluenza virus type 3: characterization and evaluation as a subunit vaccine. J Infect Dis. 1985 Dec;152(6):1219–1230. doi: 10.1093/infdis/152.6.1219. [DOI] [PubMed] [Google Scholar]
  32. Ray R., Compans R. W. Glycoproteins of human parainfluenza virus type 3: affinity purification, antigenic characterization and reconstitution into lipid vesicles. J Gen Virol. 1987 Feb;68(Pt 2):409–418. doi: 10.1099/0022-1317-68-2-409. [DOI] [PubMed] [Google Scholar]
  33. Ray R., Compans R. W. Monoclonal antibodies reveal extensive antigenic differences between the hemagglutinin-neuraminidase glycoproteins of human and bovine parainfluenza 3 viruses. Virology. 1986 Jan 15;148(1):232–236. doi: 10.1016/0042-6822(86)90420-4. [DOI] [PubMed] [Google Scholar]
  34. Reitter J. N., Sergel T., Morrison T. G. Mutational analysis of the leucine zipper motif in the Newcastle disease virus fusion protein. J Virol. 1995 Oct;69(10):5995–6004. doi: 10.1128/jvi.69.10.5995-6004.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sanderson C. M., Wu H. H., Nayak D. P. Sendai virus M protein binds independently to either the F or the HN glycoprotein in vivo. J Virol. 1994 Jan;68(1):69–76. doi: 10.1128/jvi.68.1.69-76.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Scheid A., Choppin P. W. Identification of biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis, and infectivity of proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology. 1974 Feb;57(2):475–490. doi: 10.1016/0042-6822(74)90187-1. [DOI] [PubMed] [Google Scholar]
  37. Sergel-Germano T., McQuain C., Morrison T. Mutations in the fusion peptide and heptad repeat regions of the Newcastle disease virus fusion protein block fusion. J Virol. 1994 Nov;68(11):7654–7658. doi: 10.1128/jvi.68.11.7654-7658.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sergel T., McGinnes L. W., Peeples M. E., Morrison T. G. The attachment function of the Newcastle disease virus hemagglutinin-neuraminidase protein can be separated from fusion promotion by mutation. Virology. 1993 Apr;193(2):717–726. doi: 10.1006/viro.1993.1180. [DOI] [PubMed] [Google Scholar]
  39. Tanabayashi K., Compans R. W. Functional interaction of paramyxovirus glycoproteins: identification of a domain in Sendai virus HN which promotes cell fusion. J Virol. 1996 Sep;70(9):6112–6118. doi: 10.1128/jvi.70.9.6112-6118.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tanabayashi K., Takeuchi K., Okazaki K., Hishiyama M., Yamada A. Expression of mumps virus glycoproteins in mammalian cells from cloned cDNAs: both F and HN proteins are required for cell fusion. Virology. 1992 Apr;187(2):801–804. doi: 10.1016/0042-6822(92)90482-5. [DOI] [PubMed] [Google Scholar]
  41. Tanaka Y., Heminway B. R., Galinski M. S. Down-regulation of paramyxovirus hemagglutinin-neuraminidase glycoprotein surface expression by a mutant fusion protein containing a retention signal for the endoplasmic reticulum. J Virol. 1996 Aug;70(8):5005–5015. doi: 10.1128/jvi.70.8.5005-5015.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Taylor J., Pincus S., Tartaglia J., Richardson C., Alkhatib G., Briedis D., Appel M., Norton E., Paoletti E. Vaccinia virus recombinants expressing either the measles virus fusion or hemagglutinin glycoprotein protect dogs against canine distemper virus challenge. J Virol. 1991 Aug;65(8):4263–4274. doi: 10.1128/jvi.65.8.4263-4274.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tsurudome M., Kawano M., Yuasa T., Tabata N., Nishio M., Komada H., Ito Y. Identification of regions on the hemagglutinin-neuraminidase protein of human parainfluenza virus type 2 important for promoting cell fusion. Virology. 1995 Oct 20;213(1):190–203. doi: 10.1006/viro.1995.1559. [DOI] [PubMed] [Google Scholar]
  44. White J. M. Membrane fusion. Science. 1992 Nov 6;258(5084):917–924. doi: 10.1126/science.1439803. [DOI] [PubMed] [Google Scholar]
  45. Wild T. F., Fayolle J., Beauverger P., Buckland R. Measles virus fusion: role of the cysteine-rich region of the fusion glycoprotein. J Virol. 1994 Nov;68(11):7546–7548. doi: 10.1128/jvi.68.11.7546-7548.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wild T. F., Malvoisin E., Buckland R. Measles virus: both the haemagglutinin and fusion glycoproteins are required for fusion. J Gen Virol. 1991 Feb;72(Pt 2):439–442. doi: 10.1099/0022-1317-72-2-439. [DOI] [PubMed] [Google Scholar]
  47. Yao Q., Compans R. W. Differences in the role of the cytoplasmic domain of human parainfluenza virus fusion proteins. J Virol. 1995 Nov;69(11):7045–7053. doi: 10.1128/jvi.69.11.7045-7053.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yao Q., Compans R. W. Peptides corresponding to the heptad repeat sequence of human parainfluenza virus fusion protein are potent inhibitors of virus infection. Virology. 1996 Sep 1;223(1):103–112. doi: 10.1006/viro.1996.0459. [DOI] [PubMed] [Google Scholar]

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