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. 1989 Feb;63(2):833–840. doi: 10.1128/jvi.63.2.833-840.1989

Characterization of bovine respiratory syncytial virus proteins and mRNAs and generation of cDNA clones to the viral mRNAs.

R A Lerch 1, E J Stott 1, G W Wertz 1
PMCID: PMC247757  PMID: 2911122

Abstract

We have characterized the proteins and mRNAs of bovine respiratory syncytial (BRS) virus strain 391-2 and constructed cDNA clones corresponding to 9 of the 10 BRS virus mRNAs. The proteins of BRS virus-infected cells were compared with the proteins from human respiratory syncytial (HRS) virus-infected cells. Nine proteins specific to BRS virus-infected cells, corresponding to nine HRS virus proteins, were identified. Only a BRS virus polymerase protein remains to be identified. The BRS virus G glycoprotein showed major antigenic differences from the HRS virus G glycoprotein by immunoprecipitation and Western (immuno-) blot analysis, whereas the BRS virus F, N, M, and P proteins showed antigenic cross-reactivity with their HRS virus counterparts. Analysis of RNAs from BRS virus-infected cells showed virus-specific RNAs which had electrophoretic mobilities similar to those of mRNAs of HRS virus but which hybridized poorly or not at all with HRS virus-specific probes in Northern (RNA) blot analysis. To analyze the BRS virus RNAs further, cDNA clones to the BRS virus mRNAs were generated. Nine separate groups of clones were identified and shown to correspond to nine BRS virus mRNAs by Northern blot analysis. A 10th BRS virus large mRNA was identified by analogy with the HRS virus polymerase mRNA. These data show that like HRS virus, BRS virus has 10 genes coding for 10 mRNAs.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson L. J., Hierholzer J. C., Tsou C., Hendry R. M., Fernie B. F., Stone Y., McIntosh K. Antigenic characterization of respiratory syncytial virus strains with monoclonal antibodies. J Infect Dis. 1985 Apr;151(4):626–633. doi: 10.1093/infdis/151.4.626. [DOI] [PubMed] [Google Scholar]
  2. BLOUNT R. E., Jr, MORRIS J. A., SAVAGE R. E. Recovery of cytopathogenic agent from chimpanzees with coryza. Proc Soc Exp Biol Med. 1956 Jul;92(3):544–549. doi: 10.3181/00379727-92-22538. [DOI] [PubMed] [Google Scholar]
  3. Baker J. C., Ames T. R., Markham R. J. Serologic studies of bovine respiratory syncytial virus in Minnesota cattle. Am J Vet Res. 1985 Apr;46(4):891–892. [PubMed] [Google Scholar]
  4. Baker J. C., Werdin R. E., Ames T. R., Markham R. J., Larson V. L. Study on the etiologic role of bovine respiratory syncytial virus in pneumonia of dairy calves. J Am Vet Med Assoc. 1986 Jul 1;189(1):66–70. [PubMed] [Google Scholar]
  5. CHANOCK R., ROIZMAN B., MYERS R. Recovery from infants with respiratory illness of a virus related to chimpanzee coryza agent (CCA). I. Isolation, properties and characterization. Am J Hyg. 1957 Nov;66(3):281–290. doi: 10.1093/oxfordjournals.aje.a119901. [DOI] [PubMed] [Google Scholar]
  6. Cash P., Pringle C. R., Preston C. M. The polypeptides of human respiratory syncytial virus: products of cell-free protein synthesis and post-translational modifications. Virology. 1979 Jan 30;92(2):375–384. doi: 10.1016/0042-6822(79)90142-9. [DOI] [PubMed] [Google Scholar]
  7. Cash P., Wunner W. H., Pringle C. R. A comparison of the polypeptides of human and bovine respiratory syncytial viruses and murine pneumonia virus. Virology. 1977 Oct 15;82(2):369–379. doi: 10.1016/0042-6822(77)90012-5. [DOI] [PubMed] [Google Scholar]
  8. Castleman W. L., Chandler S. K., Slauson D. O. Experimental bovine respiratory syncytial virus infection in conventional calves: ultrastructural respiratory lesions. Am J Vet Res. 1985 Mar;46(3):554–560. [PubMed] [Google Scholar]
  9. Castleman W. L., Lay J. C., Dubovi E. J., Slauson D. O. Experimental bovine respiratory syncytial virus infection in conventional calves: light microscopic lesions, microbiology, and studies on lavaged lung cells. Am J Vet Res. 1985 Mar;46(3):547–553. [PubMed] [Google Scholar]
  10. Collins P. L., Dickens L. E., Buckler-White A., Olmsted R. A., Spriggs M. K., Camargo E., Coelingh K. V. Nucleotide sequences for the gene junctions of human respiratory syncytial virus reveal distinctive features of intergenic structure and gene order. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4594–4598. doi: 10.1073/pnas.83.13.4594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Collins P. L., Huang Y. T., Wertz G. W. Identification of a tenth mRNA of respiratory syncytial virus and assignment of polypeptides to the 10 viral genes. J Virol. 1984 Feb;49(2):572–578. doi: 10.1128/jvi.49.2.572-578.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Collins P. L., Olmsted R. A., Spriggs M. K., Johnson P. R., Buckler-White A. J. Gene overlap and site-specific attenuation of transcription of the viral polymerase L gene of human respiratory syncytial virus. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5134–5138. doi: 10.1073/pnas.84.15.5134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Davis N. L., Wertz G. W. Synthesis of vesicular stomatitis virus negative-strand RNA in vitro: dependence on viral protein synthesis. J Virol. 1982 Mar;41(3):821–832. doi: 10.1128/jvi.41.3.821-832.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Evermann J. F., Liggitt H. D., Parish S. M., Ward A. C., LeaMaster B. R. Properties of a respiratory syncytial virus isolated from a sheep with rhinitis. Am J Vet Res. 1985 Apr;46(4):947–951. [PubMed] [Google Scholar]
  16. Grunstein M., Wallis J. Colony hybridization. Methods Enzymol. 1979;68:379–389. doi: 10.1016/0076-6879(79)68027-8. [DOI] [PubMed] [Google Scholar]
  17. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  18. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  19. Huang Y. T., Collins P. L., Wertz G. W. Characterization of the 10 proteins of human respiratory syncytial virus: identification of a fourth envelope-associated protein. Virus Res. 1985 Mar;2(2):157–173. doi: 10.1016/0168-1702(85)90246-1. [DOI] [PubMed] [Google Scholar]
  20. Huang Y. T., Wertz G. W. The genome of respiratory syncytial virus is a negative-stranded RNA that codes for at least seven mRNA species. J Virol. 1982 Jul;43(1):150–157. doi: 10.1128/jvi.43.1.150-157.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kimman T. G., Westenbrink F., Schreuder B. E., Straver P. J. Local and systemic antibody response to bovine respiratory syncytial virus infection and reinfection in calves with and without maternal antibodies. J Clin Microbiol. 1987 Jun;25(6):1097–1106. doi: 10.1128/jcm.25.6.1097-1106.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lehmkuhl H. D., Smith M. H., Cutlip R. C. Morphogenesis and structure of caprine respiratory syncytial virus. Arch Virol. 1980;65(3-4):269–276. doi: 10.1007/BF01314543. [DOI] [PubMed] [Google Scholar]
  24. Levine S., Klaiber-Franco R., Paradiso P. R. Demonstration that glycoprotein G is the attachment protein of respiratory syncytial virus. J Gen Virol. 1987 Sep;68(Pt 9):2521–2524. doi: 10.1099/0022-1317-68-9-2521. [DOI] [PubMed] [Google Scholar]
  25. Matumoto M., Inaba Y., Kurogi H., Sato K., Omori T. Bovine respiratory syncytial virus: host range in laboratory animals and cell cultures. Arch Gesamte Virusforsch. 1974;44(3):280–290. doi: 10.1007/BF01240616. [DOI] [PubMed] [Google Scholar]
  26. Mufson M. A., Orvell C., Rafnar B., Norrby E. Two distinct subtypes of human respiratory syncytial virus. J Gen Virol. 1985 Oct;66(Pt 10):2111–2124. doi: 10.1099/0022-1317-66-10-2111. [DOI] [PubMed] [Google Scholar]
  27. Orvell C., Norrby E., Mufson M. A. Preparation and characterization of monoclonal antibodies directed against five structural components of human respiratory syncytial virus subgroup B. J Gen Virol. 1987 Dec;68(Pt 12):3125–3135. doi: 10.1099/0022-1317-68-12-3125. [DOI] [PubMed] [Google Scholar]
  28. Paccaud M. F., Jacquier C. A respiratory syncytial virus of bovine origin. Arch Gesamte Virusforsch. 1970;30(4):327–342. doi: 10.1007/BF01258363. [DOI] [PubMed] [Google Scholar]
  29. Pringle C. R., Cross A. Neutralisation of respiratory syncytial virus by cat serum. Nature. 1978 Nov 30;276(5687):501–502. doi: 10.1038/276501a0. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Samson A. C., Willcocks M. M., Routledge E. G., Morgan L. A., Toms G. L. A neutralizing monoclonal antibody to respiratory syncytial virus which binds to both F1 and F2 components of the fusion protein. J Gen Virol. 1986 Jul;67(Pt 7):1479–1483. doi: 10.1099/0022-1317-67-7-1479. [DOI] [PubMed] [Google Scholar]
  32. Stott E. J., Taylor G. Respiratory syncytial virus. Brief review. Arch Virol. 1985;84(1-2):1–52. doi: 10.1007/BF01310552. [DOI] [PubMed] [Google Scholar]
  33. Stott E. J., Thomas L. H., Collins A. P., Crouch S., Jebbett J., Smith G. S., Luther P. D., Caswell R. A survey of virus infections of the respiratory tract of cattle and their association with disease. J Hyg (Lond) 1980 Oct;85(2):257–270. doi: 10.1017/s0022172400063294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Verhoeff J., van Nieuwstadt A. P. BRS virus, PI3 virus and BHV1 infections of young stock on self-contained dairy farms: epidemiological and clinical findings. Vet Rec. 1984 Mar 24;114(12):288–293. doi: 10.1136/vr.114.12.288. [DOI] [PubMed] [Google Scholar]
  35. Wertz G. W., Collins P. L., Huang Y., Gruber C., Levine S., Ball L. A. Nucleotide sequence of the G protein gene of human respiratory syncytial virus reveals an unusual type of viral membrane protein. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4075–4079. doi: 10.1073/pnas.82.12.4075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wertz G. W., Davis N. L. RNase III cleaves vesicular stomatitis virus genome-length RNAs but fails to cleave viral mRNA's. J Virol. 1979 Apr;30(1):108–115. doi: 10.1128/jvi.30.1.108-115.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wertz G. W., Davis N. Characterization and mapping of RNase III cleavage sites in VSV genome RNA. Nucleic Acids Res. 1981 Dec 11;9(23):6487–6503. doi: 10.1093/nar/9.23.6487. [DOI] [PMC free article] [PubMed] [Google Scholar]

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