Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1986 Mar;57(3):1010–1015. doi: 10.1128/jvi.57.3.1010-1015.1986

Characterization and translation of transmissible gastroenteritis virus mRNAs.

L Jacobs, B A van der Zeijst, M C Horzinek
PMCID: PMC252834  PMID: 3005607

Abstract

Three protein species were identified in purified transmissible gastroenteritis virus particles (strain Purdue). They are thought to represent constituents of the peplomer (E2; molecular weights of 280,000 and 240,000), the envelope (E1; molecular weights of 28,000, 31,500, and 33,000), and the nucleocapsid (N; molecular weight of 48,000). In infected cells, proteins with molecular weights of 195,000 (E2), 48,000 (N), and 28,000 (E1) were detected. Tunicamycin, an inhibitor of N glycosylation, prevented the appearance of polypeptides with molecular weights of 195,000 and 28,000 in infected cells; instead, proteins with molecular weights of 160,000 and 25,000 were observed. One minor and five major mRNA species were detected in porcine cells after infection. Their size was determined to be 23.6 kilobases (kb) (RNA1), 8.4 kb (RNA3), 3.8 kb (RNA4), 3.0 kb (RNA5), 2.6 kb (RNA6), and 1.9 kb (RNA7). The RNAs were translated in vitro. RNA7 was shown to code for the N protein. Although complete separation of RNA6 could not be achieved, it was shown to encode an unglycosylated (molecular weight of 25,000) precursor of E1 (molecular weight of 28,000). RNA4 was translated into a nonstructural protein with a molecular weight of 24,000. Translation of RNA3 resulted in proteins with molecular weights of 250,000 and 130,000 and smaller molecules which could be precipitated with a monoclonal antibody directed against E2.

Full text

PDF
1010

Images in this article

1012Image
on p.1012
1013Image
on p.1013
1014Image
on p.1014

Selected References

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

  1. Boursnell M. E., Brown T. D., Binns M. M. Sequence of the membrane protein gene from avian coronavirus IBV. Virus Res. 1984;1(4):303–313. doi: 10.1016/0168-1702(84)90019-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boursnell M. E., Brown T. D. Sequencing of coronavirus IBV genomic RNA: a 195-base open reading frame encoded by mRNA B. Gene. 1984 Jul-Aug;29(1-2):87–92. doi: 10.1016/0378-1119(84)90169-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brian D. A., Dennis D. E., Guy J. S. Genome of porcine transmissible gastroenteritis virus. J Virol. 1980 May;34(2):410–415. doi: 10.1128/jvi.34.2.410-415.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown T. D., Boursnell M. E., Binns M. M. A leader sequence is present on mRNA A of avian infectious bronchitis virus. J Gen Virol. 1984 Aug;65(Pt 8):1437–1442. doi: 10.1099/0022-1317-65-8-1437. [DOI] [PubMed] [Google Scholar]
  5. Cheley S., Anderson R., Cupples M. J., Chan E. C., Morris V. L. Intracellular murine hepatitis virus-specific RNAs contain common sequences. Virology. 1981 Jul 30;112(2):596–604. doi: 10.1016/0042-6822(81)90305-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dennis D. E., Brian D. A. RNA-dependent RNA polymerase activity in coronavirus- infected cells. J Virol. 1982 Apr;42(1):153–164. doi: 10.1128/jvi.42.1.153-164.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Garwes D. J., Bountiff L., Millson G. C., Elleman C. J. Defective replication of porcine transmissible gastroenteritis virus in a continuous cell line. Adv Exp Med Biol. 1984;173:79–93. doi: 10.1007/978-1-4615-9373-7_7. [DOI] [PubMed] [Google Scholar]
  8. Garwes D. J., Pocock D. H., Pike B. V. Isolation of subviral components from transmissible gastroenteritis virus. J Gen Virol. 1976 Aug;32(2):283–294. doi: 10.1099/0022-1317-32-2-283. [DOI] [PubMed] [Google Scholar]
  9. Garwes D. J., Pocock D. H. The polypeptide structure of transmissible gastroenteritis virus. J Gen Virol. 1975 Oct;29(1):25–34. doi: 10.1099/0022-1317-29-1-25. [DOI] [PubMed] [Google Scholar]
  10. Holmes K. V., Doller E. W., Sturman L. S. Tunicamycin resistant glycosylation of coronavirus glycoprotein: demonstration of a novel type of viral glycoprotein. Virology. 1981 Dec;115(2):334–344. doi: 10.1016/0042-6822(81)90115-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Horzinek M. C., Lutz H., Pedersen N. C. Antigenic relationships among homologous structural polypeptides of porcine, feline, and canine coronaviruses. Infect Immun. 1982 Sep;37(3):1148–1155. doi: 10.1128/iai.37.3.1148-1155.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lai M. M., Baric R. S., Brayton P. R., Stohlman S. A. Characterization of leader RNA sequences on the virion and mRNAs of mouse hepatitis virus, a cytoplasmic RNA virus. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3626–3630. doi: 10.1073/pnas.81.12.3626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lai M. M., Brayton P. R., Armen R. C., Patton C. D., Pugh C., Stohlman S. A. Mouse hepatitis virus A59: mRNA structure and genetic localization of the sequence divergence from hepatotropic strain MHV-3. J Virol. 1981 Sep;39(3):823–834. doi: 10.1128/jvi.39.3.823-834.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leibowitz J. L., Weiss S. R., Paavola E., Bond C. W. Cell-free translation of murine coronavirus RNA. J Virol. 1982 Sep;43(3):905–913. doi: 10.1128/jvi.43.3.905-913.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leibowitz J. L., Wilhelmsen K. C., Bond C. W. The virus-specific intracellular RNA species of two murine coronaviruses: MHV-a59 and MHV-JHM. Virology. 1981 Oct 15;114(1):39–51. doi: 10.1016/0042-6822(81)90250-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Niemann H., Klenk H. D. Coronavirus glycoprotein E1, a new type of viral glycoprotein. J Mol Biol. 1981 Dec 25;153(4):993–1010. doi: 10.1016/0022-2836(81)90463-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pedersen N. C., Ward J., Mengeling W. L. Antigenic relationship of the feline infectious peritonitis virus to coronaviruses of other species. Arch Virol. 1978;58(1):45–53. doi: 10.1007/BF01315534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rottier P. J., Horzinek M. C., van der Zeijst B. A. Viral protein synthesis in mouse hepatitis virus strain A59-infected cells: effect of tunicamycin. J Virol. 1981 Nov;40(2):350–357. doi: 10.1128/jvi.40.2.350-357.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rottier P. J., Spaan W. J., Horzinek M. C., van der Zeijst B. A. Translation of three mouse hepatitis virus strain A59 subgenomic RNAs in Xenopus laevis oocytes. J Virol. 1981 Apr;38(1):20–26. doi: 10.1128/jvi.38.1.20-26.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Siddell S. G., Wege H., Barthel A., ter Meulen V. Coronavirus JHM: cell-free synthesis of structural protein p60. J Virol. 1980 Jan;33(1):10–17. doi: 10.1128/jvi.33.1.10-17.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Siddell S. Coronavirus JHM: coding assignments of subgenomic mRNAs. J Gen Virol. 1983 Jan;64(Pt 1):113–125. doi: 10.1099/0022-1317-64-1-113. [DOI] [PubMed] [Google Scholar]
  22. Siddell S., Wege H., Barthel A., ter Meulen V. Coronavirus JHM: intracellular protein synthesis. J Gen Virol. 1981 Mar;53(Pt 1):145–155. doi: 10.1099/0022-1317-53-1-145. [DOI] [PubMed] [Google Scholar]
  23. Siddell S., Wege H., ter Meulen V. The structure and replication of coronaviruses. Curr Top Microbiol Immunol. 1982;99:131–163. doi: 10.1007/978-3-642-68528-6_4. [DOI] [PubMed] [Google Scholar]
  24. Spaan W. J., Rottier P. J., Horzinek M. C., van der Zeijst B. A. Isolation and identification of virus-specific mRNAs in cells infected with mouse hepatitis virus (MHV-A59). Virology. 1981 Jan 30;108(2):424–434. doi: 10.1016/0042-6822(81)90449-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Spaan W., Delius H., Skinner M. A., Armstrong J., Rottier P., Smeekens S., Siddell S. G., van der Zeijst B. Transcription strategy of coronaviruses: fusion of non-contiguous sequences during mRNA synthesis. Adv Exp Med Biol. 1984;173:173–186. doi: 10.1007/978-1-4615-9373-7_18. [DOI] [PubMed] [Google Scholar]
  26. Spaan W., Delius H., Skinner M., Armstrong J., Rottier P., Smeekens S., van der Zeijst B. A., Siddell S. G. Coronavirus mRNA synthesis involves fusion of non-contiguous sequences. EMBO J. 1983;2(10):1839–1844. doi: 10.1002/j.1460-2075.1983.tb01667.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stern D. F., Sefton B. M. Coronavirus multiplication: locations of genes for virion proteins on the avian infectious bronchitis virus genome. J Virol. 1984 Apr;50(1):22–29. doi: 10.1128/jvi.50.1.22-29.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stern D. F., Sefton B. M. Coronavirus proteins: biogenesis of avian infectious bronchitis virus virion proteins. J Virol. 1982 Dec;44(3):794–803. doi: 10.1128/jvi.44.3.794-803.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stern D. F., Sefton B. M. Coronavirus proteins: structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins. J Virol. 1982 Dec;44(3):804–812. doi: 10.1128/jvi.44.3.804-812.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Struck D. K., Lennarz W. J. Evidence for the participation of saccharide-lipids in the synthesis of the oligosaccharide chain of ovalbumin. J Biol Chem. 1977 Feb 10;252(3):1007–1013. [PubMed] [Google Scholar]
  31. Sturman L. S., Holmes K. V. Proteolytic cleavage of peplomeric glycoprotein E2 of MHV yields two 90K subunits and activates cell fusion. Adv Exp Med Biol. 1984;173:25–35. doi: 10.1007/978-1-4615-9373-7_3. [DOI] [PubMed] [Google Scholar]
  32. Sturman L. S., Holmes K. V. The molecular biology of coronaviruses. Adv Virus Res. 1983;28:35–112. doi: 10.1016/S0065-3527(08)60721-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tajima M. Morphology of transmissible gastroenteritis virus of pigs. A possible member of coronaviruses. Brief report. Arch Gesamte Virusforsch. 1970;29(1):105–108. doi: 10.1007/BF01253886. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES