Abstract
cDNA clones representing five of the genes of Sendai virus (P, HN, NP, F, and M) were isolated and used to identify the viral mRNAs by hybridization. Five mRNAs that were monocistronic transcripts of these genes were identified. A sixth transcript, which was identified on the basis of size and of hybridization to viral RNA but not to the cDNA of the other five genes, is thought to represent the message for the L protein. In addition, polycistronic transcripts of the NP and P genes and of the M and F genes were also found. The latter establishes the position of the F gene adjacent to the M gene; these results confirm and extend the previously reported partial gene order of the virus. Nucleotide sequences and derived amino acid sequences of two biologically important regions of the F protein--approximately 25% of F proximal to its COOH terminus and the region spanning the site of the proteolytic cleavage that activates the fusion activity of the protein--are presented. The F protein has an unusually large "cytoplasmic domain" of 42 amino acids beyond the hydrophobic region by which it is anchored in the viral membrane. A single possible trypsin cleavage site was found at the junction of the F1 and F2 polypeptides, and 26 hydrophobic amino acids extend from this cleavage site at the NH2 terminus of the F1 polypeptide.
Full text
PDF![7732](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab76/392226/32fb2a1074b5/pnas00625-0071.png)
![7733](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab76/392226/67d991702b89/pnas00625-0072.png)
![7734](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab76/392226/388ea499d28b/pnas00625-0073.png)
![7735](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab76/392226/aef1c71a4f7d/pnas00625-0074.png)
![7736](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab76/392226/071676a2b3f9/pnas00625-0075.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Asano K., Murachi T., Asano A. Structural requirements for hemolytic activity of F-glycoprotein of HVJ (Sendai virus) studied by proteolytic dissection. J Biochem. 1983 Mar;93(3):733–741. doi: 10.1093/jb/93.3.733. [DOI] [PubMed] [Google Scholar]
- Bosch F. X., Garten W., Klenk H. D., Rott R. Proteolytic cleavage of influenza virus hemagglutinins: primary structure of the connecting peptide between HA1 and HA2 determines proteolytic cleavability and pathogenicity of Avian influenza viruses. Virology. 1981 Sep;113(2):725–735. doi: 10.1016/0042-6822(81)90201-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Collins P. L., Huang Y. T., Wertz G. W. Nucleotide sequence of the gene encoding the fusion (F) glycoprotein of human respiratory syncytial virus. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7683–7687. doi: 10.1073/pnas.81.24.7683. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collins P. L., Wertz G. W., Ball L. A., Hightower L. E. Coding assignments of the five smaller mRNAs of Newcastle disease virus. J Virol. 1982 Sep;43(3):1024–1031. doi: 10.1128/jvi.43.3.1024-1031.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collins P. L., Wertz G. W. cDNA cloning and transcriptional mapping of nine polyadenylylated RNAs encoded by the genome of human respiratory syncytial virus. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3208–3212. doi: 10.1073/pnas.80.11.3208. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dowling P. C., Giorgi C., Roux L., Dethlefsen L. A., Galantowicz M. E., Blumberg B. M., Kolakofsky D. Molecular cloning of the 3'-proximal third of Sendai virus genome. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5213–5216. doi: 10.1073/pnas.80.17.5213. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Etkind P. R., Cross R. K., Lamb R. A., Merz D. C., Choppin P. W. In vitro synthesis of structural and nonstructural proteins of Sendai and SV5 viruses. Virology. 1980 Jan 15;100(1):22–33. doi: 10.1016/0042-6822(80)90548-6. [DOI] [PubMed] [Google Scholar]
- Gallione C. J., Rose J. K. Nucleotide sequence of a cDNA clone encoding the entire glycoprotein from the New Jersey serotype of vesicular stomatitis virus. J Virol. 1983 Apr;46(1):162–169. doi: 10.1128/jvi.46.1.162-169.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garten W., Bosch F. X., Linder D., Rott R., Klenk H. D. Proteolytic activation of the influenza virus hemagglutinin: The structure of the cleavage site and the enzymes involved in cleavage. Virology. 1981 Dec;115(2):361–374. doi: 10.1016/0042-6822(81)90117-3. [DOI] [PubMed] [Google Scholar]
- Gething M. J., White J. M., Waterfield M. D. Purification of the fusion protein of Sendai virus: analysis of the NH2-terminal sequence generated during precursor activation. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2737–2740. doi: 10.1073/pnas.75.6.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giorgi C., Blumberg B. M., Kolakofsky D. Sendai virus contains overlapping genes expressed from a single mRNA. Cell. 1983 Dec;35(3 Pt 2):829–836. doi: 10.1016/0092-8674(83)90115-0. [DOI] [PubMed] [Google Scholar]
- Glazier K., Raghow R., Kingsbury D. W. Regulation of Sendai virus transcription: evidence for a single promoter in vivo. J Virol. 1977 Mar;21(3):863–871. doi: 10.1128/jvi.21.3.863-871.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gupta K. C., Kingsbury D. W. Conserved polyadenylation signals in two negative-strand RNA virus families. Virology. 1982 Jul 30;120(2):518–523. doi: 10.1016/0042-6822(82)90055-1. [DOI] [PubMed] [Google Scholar]
- Gupta K. C., Morgan E. M., Kitchingman G., Kingsbury D. W. Molecular clones representing Sendai virus genes P, NP and M. J Gen Virol. 1983 Aug;64(Pt 8):1679–1688. doi: 10.1099/0022-1317-64-8-1679. [DOI] [PubMed] [Google Scholar]
- Herman R. C., Schubert M., Keene J. D., Lazzarini R. A. Polycistronic vesicular stomatitis virus RNA transcripts. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4662–4665. doi: 10.1073/pnas.77.8.4662. [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. Enhancement of membrane-fusing activity of sendai virus by exposure of the virus to basic pH is correlated with a conformational change in the fusion protein. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5862–5866. doi: 10.1073/pnas.79.19.5862. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsu M. C., Scheid A., Choppin P. W. Fusion of Sendai virus with liposomes: dependence on the viral fusion protein (F) and the lipid composition of liposomes. Virology. 1983 Apr 15;126(1):361–369. doi: 10.1016/0042-6822(83)90485-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Huddleston J. A., Brownlee G. G. The sequence of the nucleoprotein gene of human influenza A virus, strain A/NT/60/68. Nucleic Acids Res. 1982 Feb 11;10(3):1029–1038. doi: 10.1093/nar/10.3.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inglis S. C., Mahy B. W. Polypeptides specified by the influenza virus genome. 3. Control of synthesis in infected cells. Virology. 1979 May;95(1):154–164. doi: 10.1016/0042-6822(79)90410-0. [DOI] [PubMed] [Google Scholar]
- Klenk H. D., Rott R., Orlich M., Blödorn J. Activation of influenza A viruses by trypsin treatment. Virology. 1975 Dec;68(2):426–439. doi: 10.1016/0042-6822(75)90284-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Lamb R. A., Choppin P. W., Chanock R. M., Lai C. J. Mapping of the two overlapping genes for polypeptides NS1 and NS2 on RNA segment 8 of influenza virus genome. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1857–1861. doi: 10.1073/pnas.77.4.1857. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lamb R. A., Choppin P. W. Segment 8 of the influenza virus genome is unique in coding for two polypeptides. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4908–4912. doi: 10.1073/pnas.76.10.4908. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lamb R. A., Etkind P. R., Choppin P. W. Evidence for a ninth influenza viral polypeptide. Virology. 1978 Nov;91(1):60–78. doi: 10.1016/0042-6822(78)90355-0. [DOI] [PubMed] [Google Scholar]
- Lazarowitz S. G., Choppin P. W. Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide. Virology. 1975 Dec;68(2):440–454. doi: 10.1016/0042-6822(75)90285-8. [DOI] [PubMed] [Google Scholar]
- Lazarowitz S. G., Compans R. W., Choppin P. W. Influenza virus structural and nonstructural proteins in infected cells and their plasma membranes. Virology. 1971 Dec;46(3):830–843. doi: 10.1016/0042-6822(71)90084-5. [DOI] [PubMed] [Google Scholar]
- Lyles D. S. Glycoproteins of Sendai virus are transmembrane proteins. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5621–5625. doi: 10.1073/pnas.76.11.5621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Masters P. S., Samuel C. E. Detection of in vivo synthesis of polycistronic mRNAs of vesicular stomatitis virus. Virology. 1984 Apr 30;134(2):277–286. doi: 10.1016/0042-6822(84)90297-6. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Morgan E. M., Re G. G., Kingsbury D. W. Complete sequence of the Sendai virus NP gene from a cloned insert. Virology. 1984 May;135(1):279–287. doi: 10.1016/0042-6822(84)90137-5. [DOI] [PubMed] [Google Scholar]
- Mountcastle W. E., Compans R. W., Choppin P. W. Proteins and glycoproteins of paramyxoviruses: a comparison of simian virus 5, Newcastle disease virus, and Sendai virus. J Virol. 1971 Jan;7(1):47–52. doi: 10.1128/jvi.7.1.47-52.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Richardson C. D., Choppin P. W. Oligopeptides that specifically inhibit membrane fusion by paramyxoviruses: studies on the site of action. Virology. 1983 Dec;131(2):518–532. doi: 10.1016/0042-6822(83)90517-2. [DOI] [PubMed] [Google Scholar]
- Richardson C. D., Scheid A., Choppin P. W. Specific inhibition of paramyxovirus and myxovirus replication by oligopeptides with amino acid sequences similar to those at the N-termini of the F1 or HA2 viral polypeptides. Virology. 1980 Aug;105(1):205–222. doi: 10.1016/0042-6822(80)90168-3. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Rose J. K. Complete intergenic and flanking gene sequences from the genome of vesicular stomatitis virus. Cell. 1980 Feb;19(2):415–421. doi: 10.1016/0092-8674(80)90515-2. [DOI] [PubMed] [Google Scholar]
- Rose J. K., Gallione C. J. Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus G and M proteins determined from cDNA clones containing the complete coding regions. J Virol. 1981 Aug;39(2):519–528. doi: 10.1128/jvi.39.2.519-528.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scheid A., Caliguiri L. A., Compans R. W., Choppin P. W. Isolation of paramyxovirus glycoproteins. Association of both hemagglutinating and neuraminidase activities with the larger SV5 glycoprotein. Virology. 1972 Dec;50(3):640–652. doi: 10.1016/0042-6822(72)90418-7. [DOI] [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. Protease activation mutants of sendai virus. Activation of biological properties by specific proteases. Virology. 1976 Jan;69(1):265–277. doi: 10.1016/0042-6822(76)90213-0. [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]
- Schubert M., Keene J. D., Herman R. C., Lazzarini R. A. Site on the vesicular stomatitis virus genome specifying polyadenylation and the end of the L gene mRNA. J Virol. 1980 May;34(2):550–559. doi: 10.1128/jvi.34.2.550-559.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shioda T., Hidaka Y., Kanda T., Shibuta H., Nomoto A., Iwasaki K. Sequence of 3,687 nucleotides from the 3' end of Sendai virus genome RNA and the predicted amino acid sequences of viral NP, P and C proteins. Nucleic Acids Res. 1983 Nov 11;11(21):7317–7330. doi: 10.1093/nar/11.21.7317. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Tozawa H., Watanabe M., Ishida N. Structural components of Sendai virus. Serological and physicochemical characterization of hemagglutinin subunit associated with neuraminidase activity. Virology. 1973 Sep;55(1):242–253. doi: 10.1016/s0042-6822(73)81027-x. [DOI] [PubMed] [Google Scholar]
- Varich N. L., Lukashevich I. S., Kaverin N. V. Newcastle disease virus-specific RNA: an analysis of 24 S and 35 S RNA transcripts. Acta Virol. 1979 Jul;23(4):273–283. [PubMed] [Google Scholar]