Abstract
The cDNA derived from the fusion gene of the virulent AV strain of Newcastle disease virus (NDV) was expressed in chicken embryo cells by using a retrovirus vector. The fusion protein expressed in this system was transported to the cell surface and was efficiently cleaved into the disulfide-linked F1-F2 form found in infectious virions. The cells expressing the fusion gene grew normally and could be passaged many times. Monolayers of these cells would plaque, in the absence of trypsin, avirulent NDV strains (strains which encode a fusion protein which is not cleaved in tissue culture). Fusion protein-expressing cells would not fuse if mixed with uninfected cells or uninfected cells expressing the hemagglutinin-neuraminidase (HN) protein. However, the fusion protein-expressing cells, if infected with avirulent strains of NDV, would fuse with uninfected cells, suggesting that fusion requires both the fusion protein and another viral protein expressed in the same cell. Fusion was also seen after transfection of the HN protein gene into fusion protein-expressing cells. Thus, the expressed fusion protein gene is capable of complementing the virus infection, providing an active cleaved fusion protein required for the spread of infection. However, the fusion protein does not mediate cell fusion unless the cell also expresses the HN protein. Fusion protein-expressing cells would not plaque influenza virus in the absence of trypsin, nor would influenza virus-infected fusion protein-expressing cells fuse with uninfected cells. Thus, the influenza virus HA protein will not substitute for the NDV HN protein in cell-to-cell fusion.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bratt M. A., Gallaher W. R. Preliminary analysis of the requirements for fusion from within and fusion from without by Newcastle disease virus. Proc Natl Acad Sci U S A. 1969 Oct;64(2):536–543. doi: 10.1073/pnas.64.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bundo-Morita K., Gibson S., Lenard J. Estimation by radiation inactivation of the size of functional units governing Sendai and influenza virus fusion. Biochemistry. 1987 Sep 22;26(19):6223–6227. doi: 10.1021/bi00393a040. [DOI] [PubMed] [Google Scholar]
- Citovsky V., Yanai P., Loyter A. The use of circular dichroism to study conformational changes induced in Sendai virus envelope glycoproteins. A correlation with the viral fusogenic activity. J Biol Chem. 1986 Feb 15;261(5):2235–2239. [PubMed] [Google Scholar]
- Gibson S., Bundo-Morita K., Portner A., Lenard J. Fusion of a Sendai mutant deficient in HN protein (ts271) with cardiolipin liposomes. Virology. 1988 Mar;163(1):226–229. doi: 10.1016/0042-6822(88)90254-1. [DOI] [PubMed] [Google Scholar]
- Gitman A. G., Loyter A. Construction of fusogenic vesicles bearing specific antibodies. Targeting of reconstituted Sendai virus envelopes towards neuraminidase-treated human erythrocytes. J Biol Chem. 1984 Aug 10;259(15):9813–9820. [PubMed] [Google Scholar]
- Glickman R. L., Syddall R. J., Iorio R. M., Sheehan J. P., Bratt M. A. Quantitative basic residue requirements in the cleavage-activation site of the fusion glycoprotein as a determinant of virulence for Newcastle disease virus. J Virol. 1988 Jan;62(1):354–356. doi: 10.1128/jvi.62.1.354-356.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hsu M. C., Scheid A., Choppin P. W. Reconstitution of membranes with individual paramyxovirus glycoproteins and phospholipid in cholate solution. Virology. 1979 Jun;95(2):476–491. doi: 10.1016/0042-6822(79)90502-6. [DOI] [PubMed] [Google Scholar]
- Hughes S. H., Greenhouse J. J., Petropoulos C. J., Sutrave P. Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors. J Virol. 1987 Oct;61(10):3004–3012. doi: 10.1128/jvi.61.10.3004-3012.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hughes S., Kosik E. Mutagenesis of the region between env and src of the SR-A strain of Rous sarcoma virus for the purpose of constructing helper-independent vectors. Virology. 1984 Jul 15;136(1):89–99. doi: 10.1016/0042-6822(84)90250-2. [DOI] [PubMed] [Google Scholar]
- Hughes S., Mellstrom K., Kosik E., Tamanoi F., Brugge J. Mutation of a termination codon affects src initiation. Mol Cell Biol. 1984 Sep;4(9):1738–1746. doi: 10.1128/mcb.4.9.1738. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunt L. A., Brown D. W., Robinson H. L., Naeve C. W., Webster R. G. Retrovirus-expressed hemagglutinin protects against lethal influenza virus infections. J Virol. 1988 Aug;62(8):3014–3019. doi: 10.1128/jvi.62.8.3014-3019.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawai S., Nishizawa M. New procedure for DNA transfection with polycation and dimethyl sulfoxide. Mol Cell Biol. 1984 Jun;4(6):1172–1174. doi: 10.1128/mcb.4.6.1172. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Markwell M. A., Fox C. F. Protein-protein interactions within paramyxoviruses identified by native disulfide bonding or reversible chemical cross-linking. J Virol. 1980 Jan;33(1):152–166. doi: 10.1128/jvi.33.1.152-166.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGinnes L. W., Morrison T. G. Nucleotide sequence of the gene encoding the Newcastle disease virus fusion protein and comparisons of paramyxovirus fusion protein sequences. Virus Res. 1986 Sep;5(4):343–356. doi: 10.1016/0168-1702(86)90028-6. [DOI] [PubMed] [Google Scholar]
- McGinnes L. W., Semerjian A., Morrison T. Conformational changes in Newcastle disease virus fusion glycoprotein during intracellular transport. J Virol. 1985 Nov;56(2):341–348. doi: 10.1128/jvi.56.2.341-348.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miura N., Uchida T., Okada Y. HVJ (Sendai virus)-induced envelope fusion and cell fusion are blocked by monoclonal anti-HN protein antibody that does not inhibit hemagglutination activity of HVJ. Exp Cell Res. 1982 Oct;141(2):409–420. doi: 10.1016/0014-4827(82)90229-4. [DOI] [PubMed] [Google Scholar]
- Morrison T. G., McGinnes L. W. Avian cells expressing the Newcastle disease virus hemagglutinin-neuraminidase protein are resistant to Newcastle disease virus infection. Virology. 1989 Jul;171(1):10–17. doi: 10.1016/0042-6822(89)90505-9. [DOI] [PubMed] [Google Scholar]
- Morrison T. G., Peeples M. E., McGinnes L. W. Conformational change in a viral glycoprotein during maturation due to disulfide bond disruption. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1020–1024. doi: 10.1073/pnas.84.4.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morrison T., Ward L. J., Semerjian A. Intracellular processing of the Newcastle disease virus fusion glycoprotein. J Virol. 1985 Mar;53(3):851–857. doi: 10.1128/jvi.53.3.851-857.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagai Y., Klenk H. D. Activation of precursors to both glycoporteins of Newcastle disease virus by proteolytic cleavage. Virology. 1977 Mar;77(1):125–134. doi: 10.1016/0042-6822(77)90412-3. [DOI] [PubMed] [Google Scholar]
- Nagai Y., Klenk H. D., Rott R. Proteolytic cleavage of the viral glycoproteins and its significance for the virulence of Newcastle disease virus. Virology. 1976 Jul 15;72(2):494–508. doi: 10.1016/0042-6822(76)90178-1. [DOI] [PubMed] [Google Scholar]
- Nakanishi M., Uchida T., Kim J., Okada Y. Glycoproteins of Sendai virus (HVJ) have a critical ratio for fusion between virus envelopes and cell membranes. Exp Cell Res. 1982 Nov;142(1):95–101. doi: 10.1016/0014-4827(82)90413-x. [DOI] [PubMed] [Google Scholar]
- Nussbaum O., Zakai N., Loyter A. Membrane-bound antiviral antibodies as receptors for Sendai virions in receptor-depleted erythrocytes. Virology. 1984 Oct 30;138(2):185–197. doi: 10.1016/0042-6822(84)90344-1. [DOI] [PubMed] [Google Scholar]
- Olmsted R. A., Elango N., Prince G. A., Murphy B. R., Johnson P. R., Moss B., Chanock R. M., Collins P. L. Expression of the F glycoprotein of respiratory syncytial virus by a recombinant vaccinia virus: comparison of the individual contributions of the F and G glycoproteins to host immunity. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7462–7466. doi: 10.1073/pnas.83.19.7462. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ozawa M., Asano A., Okada Y. Biological activities of glycoproteins of HVJ (Sendai virus) studied by reconstitution of hybrid envelope and by concanavalin A-mediated binding: a new function of HANA protein and structural requirement of F protein in hemolysis. Virology. 1979 Nov;99(1):197–202. doi: 10.1016/0042-6822(79)90055-2. [DOI] [PubMed] [Google Scholar]
- 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]
- Portner A., Scroggs R. A., Metzger D. W. Distinct functions of antigenic sites of the HN glycoprotein of Sendai virus. Virology. 1987 May;158(1):61–68. doi: 10.1016/0042-6822(87)90238-8. [DOI] [PubMed] [Google Scholar]
- Sakai Y., Shibuta H. Syncytium formation by recombinant vaccinia viruses carrying bovine parainfluenza 3 virus envelope protein genes. J Virol. 1989 Sep;63(9):3661–3668. doi: 10.1128/jvi.63.9.3661-3668.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Scheid A., Choppin P. W. Two disulfide-linked polypeptide chains constitute the active F protein of paramyxoviruses. Virology. 1977 Jul 1;80(1):54–66. doi: 10.1016/0042-6822(77)90380-4. [DOI] [PubMed] [Google Scholar]
- Sechoy O., Philippot J. R., Bienvenue A. Preparation and characterization of F-protein vesicles isolated from Sendai virus by means of octyl glucoside. Biochim Biophys Acta. 1986 May 9;857(1):1–12. doi: 10.1016/0005-2736(86)90093-3. [DOI] [PubMed] [Google Scholar]
- Sheehan J. P., Iorio R. M., Syddall R. J., Glickman R. L., Bratt M. A. Reducing agent-sensitive dimerization of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus correlates with the presence of cysteine at residue 123. Virology. 1987 Dec;161(2):603–606. doi: 10.1016/0042-6822(87)90158-9. [DOI] [PubMed] [Google Scholar]
- Smith G. W., Hightower L. E. Identification of the P proteins and other disulfide-linked and phosphorylated proteins of Newcastle disease virus. J Virol. 1981 Jan;37(1):256–267. doi: 10.1128/jvi.37.1.256-267.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steiner D. F., Quinn P. S., Chan S. J., Marsh J., Tager H. S. Processing mechanisms in the biosynthesis of proteins. Ann N Y Acad Sci. 1980;343:1–16. doi: 10.1111/j.1749-6632.1980.tb47238.x. [DOI] [PubMed] [Google Scholar]
- Toyoda T., Sakaguchi T., Imai K., Inocencio N. M., Gotoh B., Hamaguchi M., Nagai Y. Structural comparison of the cleavage-activation site of the fusion glycoprotein between virulent and avirulent strains of Newcastle disease virus. Virology. 1987 May;158(1):242–247. doi: 10.1016/0042-6822(87)90261-3. [DOI] [PubMed] [Google Scholar]
- Webster R. G., Brown L. E., Jackson D. C. Changes in the antigenicity of the hemagglutinin molecule of H3 influenza virus at acidic pH. Virology. 1983 Apr 30;126(2):587–599. doi: 10.1016/s0042-6822(83)80015-4. [DOI] [PubMed] [Google Scholar]