Abstract
The cell fusion activity of most paramyxoviruses requires coexpression of a fusion protein (F) and a hemagglutinin-neuraminidase protein (HN) which are derived from the same virus type. To define the domain of the HN protein which interacts with the F protein in a type-specific manner a series of chimeric HN proteins between two different paramyxoviruses, Sendai virus (SN) and human parainfluenza virus type 3 (PI3), was constructed and coexpressed with the SN-F protein by using the vaccinia virus T7 RNA polymerase transient-expression system. Quantitative assays were used to evaluate cell surface expression as well as fusion-promoting activities of the chimeric HN molecules. A chimeric HN protein [SN(140)] containing 140 N-terminal amino acids derived from SN-HN and the remainder (432 amino acids) derived from PI3-HN was found to promote cell fusion with the SN-F protein. In contrast, a second chimeric HN with 137 amino acids from SN-HN at the N terminus could not promote fusion with SN-F, even though the protein was expressed on the cell surface. A construct in which the PI3-HN cytoplasmic tail and transmembrane domain were substituted for those of SN in the SN(140) chimera still maintained the ability to promote cell fusion. These results indicate that a region including only 82 amino acids in the extracellular domain, adjacent to the transmembrane domain of the SN-HN protein, is important for interaction with the SN-F protein and promotion of cell fusion.
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- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [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]
- Horton R. M., Hunt H. D., Ho S. N., Pullen J. K., Pease L. R. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989 Apr 15;77(1):61–68. doi: 10.1016/0378-1119(89)90359-4. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Jambou R. C., Elango N., Venkatesan S. Proteins associated with human parainfluenza virus type 3. J Virol. 1985 Oct;56(1):298–302. doi: 10.1128/jvi.56.1.298-302.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Matsuoka Y., Chen S. Y., Compans R. W. A signal for Golgi retention in the bunyavirus G1 glycoprotein. J Biol Chem. 1994 Sep 9;269(36):22565–22573. [PubMed] [Google Scholar]
- McGinnes L. W., Morrison T. G. The role of the individual cysteine residues in the formation of the mature, antigenic HN protein of Newcastle disease virus. Virology. 1994 May 1;200(2):470–483. doi: 10.1006/viro.1994.1210. [DOI] [PubMed] [Google Scholar]
- McGinnes L., Sergel T., Morrison T. Mutations in the transmembrane domain of the HN protein of Newcastle disease virus affect the structure and activity of the protein. Virology. 1993 Sep;196(1):101–110. doi: 10.1006/viro.1993.1458. [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., 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]
- 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]
- Nussbaum O., Broder C. C., Berger E. A. Fusogenic mechanisms of enveloped-virus glycoproteins analyzed by a novel recombinant vaccinia virus-based assay quantitating cell fusion-dependent reporter gene activation. J Virol. 1994 Sep;68(9):5411–5422. doi: 10.1128/jvi.68.9.5411-5422.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Sergel T., McGinnes L. W., Morrison T. G. The fusion promotion activity of the NDV HN protein does not correlate with neuraminidase activity. Virology. 1993 Oct;196(2):831–834. doi: 10.1006/viro.1993.1541. [DOI] [PubMed] [Google Scholar]
- 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]
- Storey D. G., Dimock K., Kang C. Y. Structural characterization of virion proteins and genomic RNA of human parainfluenza virus 3. J Virol. 1984 Dec;52(3):761–766. doi: 10.1128/jvi.52.3.761-766.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taira H., Sato T., Segawa H., Chiba M., Katsumata T., Iwasaki K. Transfection of Sendai virus F gene cDNA with mutations at its cleavage site and HN gene cDNA into COS cells induces cell fusion. Arch Virol. 1995;140(1):187–194. doi: 10.1007/BF01309734. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- Yuasa T., Kawano M., Tabata N., Nishio M., Kusagawa S., Komada H., Matsumura H., Ito Y., Tsurudome M. A cell fusion-inhibiting monoclonal antibody binds to the presumed stalk domain of the human parainfluenza type 2 virus hemagglutinin-neuraminidase protein. Virology. 1995 Feb 1;206(2):1117–1125. doi: 10.1006/viro.1995.1035. [DOI] [PubMed] [Google Scholar]