Skip to main content
Elsevier - PMC COVID-19 Collection logoLink to Elsevier - PMC COVID-19 Collection
. 2004 Feb 6;139(1):138–151. doi: 10.1016/0042-6822(84)90335-0

Analysis of genomic and intracellular viral RNAs of small plaque mutants of mouse hepatitis virus, JHM strain

Shinji Makino ∗,1, Fumihiro Taguchi ∗,2, Norio Hirano , Kosaku Fujiwara
PMCID: PMC7119021  PMID: 6093379

Abstract

The genomic RNA and intracellular RNA of mouse hepatitis virus, strain JHM (MHV-JHM) and two plaque mutants (la and 2c), which have been isolated from a persistently infected culture (JHM-CC), have been analyzed by T1-resistant oligonucleotide finger-printing. The genomic RNA of the virus population (JHM-CC virus) released from different passage levels of the same persistent infection has also been analyzed. The analysis shows the locations within the genomic and intracellular RNAs of more than 45 T1-resistant oligonucleotides and confirm earlier studies (J. L. Leibowitz, K. C. Wilhelmsen, and C. W. Bond (1981), Virology114, 39–51), showing that the six subgenomic RNAs of MHV-JHM form a 3′ coterminal nested set which extends for different lengths in a 5′ direction. The analysis also identifies in each subgenomic RNA those large T1 oligonucleotides derived from noncontiguous regions of the genome during mRNA synthesis. Two important conclusions can be reached from analysis of the mutant viruses. First, the virus population released from the persistent infection represents a fairly constant mixture of viruses, and the fluctuating emergence of variants as predominant species in the culture does not occur. Second, the data indicate that for particular intracellular RNAs of mutant viruses the sequence rearrangements occurring during subgenomic mRNA synthesis are different from those in the corresponding intracellular RNA of wild-type virus. The result may indicate a potential flexibility in the leader/body fusion process that has not been previously recognized.

References

  1. Bark R.S., Stohlman S.A., Lai M.M.C. Characterization of replicative intermediate RNA of mouse hepatitis virus: Presence of leader RNA sequences on nascent chains. J. Virol. 1983;48:633–640. doi: 10.1128/jvi.48.3.633-640.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brayton P.R., Ganges R.G., Stohlman S.A. Host cell nuclear function and murine hepatitis virus replication. J. Gen. Virol. 1981;56:457–460. doi: 10.1099/0022-1317-56-2-457. [DOI] [PubMed] [Google Scholar]
  3. Brayton P.R., Lai M.M.C., Patton C.D., Stohlman S.A. Characterization of two RNA polymerase activities induced by mouse hepatitis virus. J. Virol. 1982;42:847–853. doi: 10.1128/jvi.42.3.847-853.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheley S., Anderson R., Cupples M.J., Lee Chan E.C.M., Morris V.L. Intracellular murine hepatitis virus-specific RNAs contain common sequences. Virology. 1981;112:596–604. doi: 10.1016/0042-6822(81)90305-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Herndon R.M., Griffin D.E., McCormic U., Weiner L.P. Mouse hepatitis virus-induced recurrent demyelination. Arch. Neurol. 1975;33:32–35. doi: 10.1001/archneur.1975.00490430054008. [DOI] [PubMed] [Google Scholar]
  6. Hirano N., Goto N., Makino S., Fujiwara K. Vol. 142, Plenum; New York: 1981. Persistent infection with mouse hepatitis virus JHM strain in DBT cell culture; pp. 301–308. (Advances in Experimental Medicine and Biology). [DOI] [PubMed] [Google Scholar]
  7. Holland J.J., Grabau E.A., Jones C.L., Semler B.L. Evolution of multiple genome mutations during long-term persistent infection of vesicular stomatitis virus. Cell. 1979;16:495–504. doi: 10.1016/0092-8674(79)90024-2. [DOI] [PubMed] [Google Scholar]
  8. Holland J., Spindler K., Horodyski F., Grabau E., Nichol S., Vandepol S. Rapid evolution of RNA genomes. Science (Washington, D. C.) 1982;215:1577–1585. doi: 10.1126/science.7041255. [DOI] [PubMed] [Google Scholar]
  9. Holmes K.V., Doller E.W., Sturman L.S. Tunicamycin resistant glycosylation of a coronavirus glycoprotein: Demonstration of a novel type of viral glycoprotein. Virology. 1981;115:334–344. doi: 10.1016/0042-6822(81)90115-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jacobs L., Spaan W.J.M., Horzinek M.C., van Der Zeijst B.A.M. The synthesis of the subgenomic mRNAs of mouse hepatitis virus is initiated independently: Evidence from UV transcription mapping. J. Virol. 1981;39:401–406. doi: 10.1128/jvi.39.2.401-406.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Knobler R.L., Dubois-Dalcq M., Haspel M.V., Claysmith A.P., Lampert P.W., Oldstone M.B.A. Selective localization of wild type and mutant mouse hepatitis virus (JHM strain) antigens in CNS tissue by fluorescence, light and electron microscopy. J. Neuroimmunol. 1981;1:81–92. doi: 10.1016/0165-5728(81)90010-2. [DOI] [PubMed] [Google Scholar]
  12. Lai M.M.C., 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;39:823–834. doi: 10.1128/jvi.39.3.823-834.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lai M.M.C., Fleming J.0., Stohlman S.A., Fujiwara K. Genetic heterogeneity of murine coronaviruses. Arch. Virol. 1983;78:167–176. doi: 10.1007/BF01311312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lai M.M.C., Patton C.D., Bark R.S., Stohlman S.A. Presence of leader sequences in the mRNA of mouse hepatitis virus. J. Virol. 1983;46:1027–1033. doi: 10.1128/jvi.46.3.1027-1033.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lai M.M.C., Patton C.D., Stohlman S.A. Further characterization of mRNA's of mouse hepatitis virus: Presence of common 5′-end nucleotides. J. Virol. 1982;41:557–565. doi: 10.1128/jvi.41.2.557-565.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lai M.M.C., Stohlman S.A. RNA of mouse hepatitis virus. J. Virol. 1978;26:236–242. doi: 10.1128/jvi.26.2.236-242.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Langridge L., Langridge P., Bergquist P.L. Extraction of nucleic acids from agarose gels. Anal. Biochem. 1980;103:264–271. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
  18. Leibowitz J.L., Wilhelmsen K.C., Bond C.W. The virus-specific intracellular RNA species of two murine coronavirus: MHV-A59 and MHV-JHM. Virology. 1981;114:39–51. doi: 10.1016/0042-6822(81)90250-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Leibowitz J.L., Weiss S.R., Paavola E., Bond C.W. Cell-free translation of murine coronavirus RNA. J. Virol. 1982;43:905–913. doi: 10.1128/jvi.43.3.905-913.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mahy B.W.J., Siddell S., Wege H., ter Meulen V. RNA-dependent RNA polymerase activity in murine coronavirus-infected cells. J. Gen. Virol. 1983;64:103–111. doi: 10.1099/0022-1317-64-1-103. [DOI] [PubMed] [Google Scholar]
  21. Making S., Taguchi F., Fujiwara K. Defective interfering particles of mouse hepatitis virus. Virology. 1984;133:9–17. doi: 10.1016/0042-6822(84)90420-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Making S., Taguchi F., Fujiwara K., Hayami M. Heterologous response of antiserumtreated cell clones from a persistently infected DBT cell line to mouse hepatitis virus. Japan. J. Exp. Med. 1982;52:297–302. [PubMed] [Google Scholar]
  23. Making S, Taguchi F., Hayami M., Fujiwara K. Characterization of small plaque mutants of mouse hepatitis virus, JHM strain. Microbiol. Immunol. 1983;27:445–454. doi: 10.1111/j.1348-0421.1983.tb00603.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McMaster G.K., Carmicheal G.G. Vol. 74. 1977. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange; pp. 4835–4838. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Meinkoth J., Kennedy S.I.T. Semliki Forest virus persistence in mouse L929 cells. Virology. 1980;100:141–155. doi: 10.1016/0042-6822(80)90560-7. [DOI] [PubMed] [Google Scholar]
  26. Nagashima K., Wege H., Meyermann P., Ter Meulen V. Coronavirus induced subacute demyelinating encephalitis in rats: A morphological analysis. Acts Neuropathol. (Berlin) 1978;44:63–70. doi: 10.1007/BF00691641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Niemann H., Klenk H.D. Coronavirus glycoprotein El, a new type of viral glycoprotein. J. Mol. Biol. 1981;153:993–1010. doi: 10.1016/0022-2836(81)90463-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rottier P.J.M., Horzinek M.C., van Der Zeijst B.A.M. Viral protein synthesis in mouse hepatitis virus strain A59-infected cells: Effect of tunicamycin. J. Virol. 1981;40:350–357. doi: 10.1128/jvi.40.2.350-357.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rottier P.J.M., Spaan W.J.M., Horzinek M.C., van Der Zeijst B.A.M. Translation of three mouse hepatitis virus strain A59 subgenomic RNAs in Xenopus laevis Oocytes. J. Virol. 1981;38:20–26. doi: 10.1128/jvi.38.1.20-26.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Siddell S.G. Coronavirus JHM: Tryptic peptide fingerprinting of virion proteins and intracellular polypeptides. J. Gen. Virol. 1982;62:259–269. doi: 10.1099/0022-1317-62-2-259. [DOI] [PubMed] [Google Scholar]
  31. Siddell S.G. Coronavirus JHM: Coding assignments of subgenomic mRNAs. J. Gen. Virol. 1983;64:113–125. doi: 10.1099/0022-1317-64-1-113. [DOI] [PubMed] [Google Scholar]
  32. Siddell S.G., Wege H., Barthel A., Ter Meulen V. Coronavirus JHM: Cell-free synthesis of structural protein P60. J. Virol. 1980;33:10–17. doi: 10.1128/jvi.33.1.10-17.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Siddell S.G., Wege H., ter Meulen V. The biology of coronaviruses. J. Gen. Virol. 1983;64:761–776. doi: 10.1099/0022-1317-64-4-761. [DOI] [PubMed] [Google Scholar]
  34. Spaan W., Delius H., Skinner M., Armstrong J., Rottier P., Smeekens S., van Der Zeijst B.A.M., Siddell S.G. Coronavirus mRNA synthesis involves fusion of noncontiguous sequences. EMBO J. 1983;2:1939–1944. doi: 10.1002/j.1460-2075.1983.tb01667.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Spaan W.J.M., Rottier P.J.M., Horzinek M.C., van Der Zeijst B.A.M. Isolation and identification of virus-specific mRNAs in cells infected with mouse hepatitis virus (MHV-A59) Virology. 1981;108:424–434. doi: 10.1016/0042-6822(81)90449-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Spaan W.J.M., Rottier P.J.M., Horzinek M.C., van Der Zeijst B.A.M. Sequence relationships between the genome and the intracellular RNA species 1, 3, 6 and 7 of mouse hepatitis virus strain A59. J. Virol. 1982;42:432–439. doi: 10.1128/jvi.42.2.432-439.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stohlman S.A., Brayton P.R., Fleming J.O., Weiner L.P., Lai M.M.C. Murine coronaviruses: Isolation and characterization of two plaque morphology variants of the JHM neurotropic strain. J. Gen. Virol. 1982;63:265–275. doi: 10.1099/0022-1317-63-2-265. [DOI] [PubMed] [Google Scholar]
  38. Wege H., Müller A., ter Meulen V. Genomic RNA of the murine coronavirus JHM. J. Gen. Virol. 1978;41:217–227. doi: 10.1099/0022-1317-41-2-217. [DOI] [PubMed] [Google Scholar]
  39. Wege H., Siddell S., Sturm M., ter Meulen V. Coronavirus JHM: Characterization of intracellular viral RNA. J. Gen. Virol. 1981;54:213–217. doi: 10.1099/0022-1317-54-1-213. [DOI] [PubMed] [Google Scholar]
  40. Wilhelmsen K.C., Leibowitz J.L., Bond C.W., Robb J.A. The replication of murine coronaviruses in enucleated cells. Virology. 1981;110:225–230. doi: 10.1016/0042-6822(81)90027-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yogo Y., Hirano N., Hind S., Shibuta H., Matumoto M. Polyadenylate in the virion RNA of mouse hepatitis virus. J. Biochem. 1977;82:1103–1108. doi: 10.1093/oxfordjournals.jbchem.a131782. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Virology are provided here courtesy of Elsevier

RESOURCES