Abstract
After six to eight serial undiluted passages of mouse hepatitis virus (JHM strain) in DBT cell culture, a decrease in the yield of infectious virus occurred, and with further passages fluctuating yields of infectious virus were observed. The serially passaged virus interfered with the multiplication of the standard JHM virus, but not with vesicular stomatitis virus. After sucrose equilibrium centrifugation of high passage virus, a single peak contained both infectious virus and interfering activity. This virus population resembled the original JHM virus in its structural proteins, but it contained an increased proportion of a protein with a molecular weight of 65 × 103. Genomic RNA from standard JHM virus contained a single species of RNA with a molecular weight of 5.4 × 106. After five undiluted passages, however, the virion population contained two RNA species with molecular weights of 5.4 × 106 and 5.2 × 106. RNase T1 resistant oligonucleotide finger-printing of these RNAs showed that the lower molecular weight RNA had lost several oligonucleotide spots that were present in the genomic RNA of the standard JHM virus. After several serial diluted passages of passage 10 virus, a single virus population was obtained which again had only standard virus RNA with a molecular weight of 5.4 × 106 and lacked interfering activity. These results indicated that defective interfering particles were generated by serial undiluted passages of JHM virus.
References
- Bailey O.T., Pappenheimer A.M., Cheever F.S., Daniels J.B. A murine virus (JHM) causing disseminated encephalomyelitis with extensive destruction of myelin. II. Pathology. J. Exp. Med. 1949;90:195–212. doi: 10.1084/jem.90.3.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bruton C.J., Kennedy S.I.T. Defective interfering particles of Semliki Forest virus: Structural differences between standard virus and defective-interfering particles. J. Gen. Virol. 1976;31:383–395. doi: 10.1099/0022-1317-31-3-383. [DOI] [PubMed] [Google Scholar]
- Chamberlain J.P. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal. Biochem. 1979;98:132–135. doi: 10.1016/0003-2697(79)90716-4. [DOI] [PubMed] [Google Scholar]
- Cheever F.S., Daniels J.B., Pappenheimer A.M., Bailey O.T. A murine virus (JHM) causing disseminated encephalomyelitis with extensive destruction of myelin. I. Isolation and biological properties of the virus. J. Exp. Med. 1949;90:181–194. doi: 10.1084/jem.90.3.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Wachter R., Fiers W. Preparative two-dimensional polyacrylamide gel electrophoresis of 32P-labeled RNA. Anal. Biochem. 1972;49:184–197. doi: 10.1016/0003-2697(72)90257-6. [DOI] [PubMed] [Google Scholar]
- Friedman R.M., Ramseur J.M. Mechanisms of persistent infection by cytopathic viruses in tissue culture. Arch. Virol. 1979;60:83–103. doi: 10.1007/BF01348025. [DOI] [PubMed] [Google Scholar]
- Hirano N., Fujiwara K., Hino S., Matumoto M. Replication and plaque formation of mouse hepatitis virus (MHV-2) in mouse cell line DBT culture. Arch. Gesamte Virusforsch. 1974;44:298–302. doi: 10.1007/BF01240618. [DOI] [PubMed] [Google Scholar]
- Hirano N., Goto N., Makino S., Fujiwara K. Persistent infection with mouse hepatitis virus JHM strain in DBT cell culture. In: ter Meulen V., Siddell S., Wege H., editors. The Biochemistry and Biology of Coronaviruses. Plenum; New York: 1981. pp. 301–308. [DOI] [PubMed] [Google Scholar]
- Holland J.J., Kennedy S.I.T., Semler B.L., Jones C.L., Roux L., Grabau E.A. Defective interfering RNA viruses and the host-cell response. In: Fraenkel-Conrat H., Wagner R.R., editors. Vol. 16. Plenum; New York: 1980. pp. 137–192. (Comprehensive Virology). [Google Scholar]
- Holland J.J., Villarreal L.P. Purification of defective interfering T particles of vesicular stomatitis and rabies viruses generated in vivo in brains of newborn mice. Virology. 1975;67:438–449. doi: 10.1016/0042-6822(75)90445-6. [DOI] [PubMed] [Google Scholar]
- Huang A.S., Baltimore D. Defective interfering animal viruses. In: Fraenkel-Conrat H., Wagner R.R., editors. Vol. 10. Plenum; New York: 1977. pp. 73–116. (Comprehensive Virology). [Google Scholar]
- Jacobson S., Pfau C.J. Viral pathogenesis and resistance to defective interfering particles. Nature (London) 1980;283:311–313. doi: 10.1038/283311a0. [DOI] [PubMed] [Google Scholar]
- Kumanishi T. Brain tumors induced with Rous sarcoma virus, Schmidt-Ruppin strain. 1. Induction of brain tumors in adult mice with Rous chicken sarcoma cells. Japan. J. Exp. Med. 1967;37:461–474. [PubMed] [Google Scholar]
- Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 1970;227:680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- 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 diverence 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]
- 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]
- Lai M.M.C., Stohlman S.A. Comparative analysis of RNA genomes of mouse hepatitis viruses. J. Virol. 1981;38:264–271. doi: 10.1128/jvi.38.2.661-670.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langridge L., Langridge P., Bergquist P.L. Extraction of nucleic acids from agarose gels. Anal. Biochem. 1980;103:661–670. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
- 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;114:39–51. doi: 10.1016/0042-6822(81)90250-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Makino S., Taguchi F., Fujiwara K., Hayami M. Heterologous response of antiserum-treated cell clones from a persistently infected DBT cell line to mouse hepatitis virus. Japan. J. Exp. Med. 1982;52:297–302. [PubMed] [Google Scholar]
- Makino 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]
- 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. Nat. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rima B.K., Martin S.J. Persistent infection of tissue culture cells by RNA viruses. Med. Microbiol. Immunol. 1976;162:89–118. doi: 10.1007/BF02121320. [DOI] [PubMed] [Google Scholar]
- Robb J.A., Bond C.W. Coronaviridae. In: Fraenkel-Conrat H., Wagner R.R., editors. Vol. 14. Plenum; New York: 1979. pp. 193–247. (Comprehensive Virology). [Google Scholar]
- Siddell S., Wege H., Barthel A., ter Meulen V. Coronavirus JHM: Intracellular protein synthesis. J. Gen. Virol. 1981;53:145–155. doi: 10.1099/0022-1317-53-1-145. [DOI] [PubMed] [Google Scholar]
- Siddell S., Wege H., ter Meulen V. The structure and replication of coronaviruses. In: Cooper M., Hofschneiden P.H., Koprowski H., Melschers F., Rott R., Schweiber H.G., Vogt P.K., Zinkernagel R., editors. Vol. 99. Springer-Verlag; Berlin/Heidelberg/New York: 1982. pp. 131–163. (Current Topics in Microbiology and Immunology). [Google Scholar]
- Stohlman S.A., Weiner L.P. Stability of neurotropic mouse hepatitis virus (JHM strain) during chronic infection of neuroblastoma cells. Arch. Virol. 1978;57:53–61. doi: 10.1007/BF01315637. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stohlman S.A., Brayton P.R., Fleming J.O., Weiner L.P., Lai M.M. 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]