Summary
Mouse hepatitis virus 3 (MHV-3) is highly hepatotropic in sensitive mice. Temperature-sensitive mutants (ts mutants) induced by N-methyl-N′-nitrosoguanidine and 5-fluorouracil were isolated. Twelve mutants which were able to induce the formation of syncytia at 33°C but not at the restrictive temperature (39.5°C) were selected for detailed study. No viral RNA synthesis was detected after infection at the restrictive temperature with six of the mutants (RNA−) whereas six others were RNA+, although they displayed RNA synthesis which was generally reduced. No differences have been detected in the size of the genome or the viral-intracellular RNA species found in wild type virus or ts mutant infected cells at permissive temperature. The pattern of virus-induced proteins analyzed after immunoprecipitation by SDS-PAGE was similar in wild type virus and RNA+ mutant infected cells at 39.5°C. Complementation experiments between ts mutants enabled us to distinguish five groups. Three of the groups contained RNA− mutants and two of them RNA+. Plaques made by mutants in one group displayed characteristic features that distinguished them from the wild type.
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Baric RS, Stohlman SA, Razavi MK, Lai MMC (1985) Characterization of leader-related small RNAs in coronavirus-infected cells: further evidence for leader-primed mechanism of transcription. Virus Res 3: 19–23
Bond CW, Leibowitz JL, Robb JA (1979) Pathogenic murine coronaviruses. II Characterization of virus-specific proteins of murine coronaviruses. JHMV and A 59 V. Virology 94: 371–384
Bonner WM, Laskey RA (1974) A film detection method for tritiumlabelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46: 83–88
Brayton PR, Lai MMC, Patton CD, Stohlman SA (1982) Characterization of to RNA polymerase activities induced by mouse hepatitis virus. J Virol 42: 847–853
Chelay S, Anderson R, Cupples MJ, Lee Chan ECM, Morris VL (1981) Intracellular murine hepatitis virus-specific RNAs contain common sequences. Virology 112: 596–604
Fields BN, Joklik WK (1969) Isolation and preliminary genetic and biochemical characterization of temperature-sensitive mutants of reovirus. Virology 37: 335–342
Haspel MV, Lambert PW, Oldstone MBA (1978) Temperature-sensitive mutants of mouse hepatitis virus produce a high incidence of demyelination. Proc Natl Acad Sci USA 75: 4033–4036
Hirano N, Murakami T, Taguchi F, Fujiwara K, Matumoto (1981) Comparison of mouse hepatitis strains for pathogenicity in weaning mice infected by various routes. Arch Virol 70: 69–73
Koolen MJ, Osterhaus AD, Van Steenis G, Horzinek MC, van der Zeijst BAM (1983) Temperature-sensitive mutants of mouse hepatitis virus strain A 59: isolation, characterization and neuropathogenic properties. Virology 125: 393–402
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Lai MMC, Stohlman SA (1978) The RNA of mouse hepatitis virus. J Virol 26: 236–242
Lai MMC, Brayton PR, Armen RC, Pugh C, Stohlman SA (1981) Mouse hepatitis virus A 59: mRNA structure and genetic localization of the sequence divergence from hepatotropic strain MHV-3. J Virol 39: 823–834
Lai MMC, Stohlman SA (1981) Genome structure of mouse hepatitis virus. Comparative analysis by oligonucleotide mapping. Adv Exp Med Biol 142: 69–82
Lai MMC, Patton CD, Stohlman SA (1982) Replication of mouse hepatitis virus: negative-stranded RNA and replicative from RNA are of genome length. J Virol 44: 487–492
Lamontagne LM, Dupuy JM (1984) Natural resistance of mice to mouse hepatitis virus type 3 infection is expressed in embryonic fibroblast cells. J Gen Virol 65: 1165–1171
Leibowitz JL, Wilhemsen KC, Bond CW (1981) The virus-specific intracellular RNA species of two murine coronaviruses: MHV A 59 and MHV JHM. Virology 114: 39–51
Leibowitz JL, de Vries JR, Haspel MV (1982) Genetic analysis of murine hepatitis virus strain JHM. J Virol 42: 1080–1087
Makino S, Taguchi F, Hirano N, Fujiwara K (1984) Analysis of genomic and intracellular viral RNAs of small plaque mutants of mouse hepatitis virus, JHM strain. Virology 139: 138–151
Pereira CA, Steffan AM, Kirn A (1984) Kupffer and endothelial liver cell damage renders A/J mice susceptible to mouse hepatitis virus type 3. Virus Res 1: 557–563
Robb JA, Bond CW, Leibowitz JL (1979) Pathogenic murine coronaviruses. III Biological and biochemical characterization of temperature-sensitive mutants of JHMV. Virology 94: 385–399
Rottier PJ, Horzinek MC, van der Zeijst BAM (1981) Viral protein synthesis in mouse hepatitis virus strain A 59-infected cells. Effect of tunicamycin. J Virol 40: 350–357
Siddell S, Wege H, ter Meulen V (1982) The structure and replication of coronaviruses. Curr Top Microbiol Immunol 99: 131–157
Spann WJM, Rottier PJM, Horzinek MC, van der Zeijst BAM (1981) Isolation and identification of virus-specific mRNAs in cells infected with mouse hepatitis virus (MHV-A 59). Virology 108: 424–434
Steffan AM, Pereira CA, Kirn A (1986) Role of sinusoidal cells in the course of the hepatitis induced by mouse hepatitis virus type 3 (MHV 3) in mice. In: Kirn A, Knook DL, Wisse E (eds) Cells of hepatic sinusoid, vol 1. The Kupffer Cell Foundation, Rijswijk, pp 377–378
Stern DF, Kennedy SIT (1980) Coronavirus multiplication strategy. II Mapping the avian infectious bronchitis virus intracellular RNA species to the genome. J Virol 36: 440–449
Stohlman SA, Fleming JO, Patton CD, Lai MMC (1983) Synthesis and subcellular localization of the murine coronavirus nucleocapsid protein. Virology 130: 527–532
Thomas P (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77: 5201–5203
Van der Zeijst BAM, Koolen MJ, Noten JF, Jansen EJ, Horzinek MC (1985) At least six gene products of murine coronavirus MHV-A 59 are required to make functional mRNAs. In: Koolen MJ: Murine coronavirus and demyelination. Thesis, University of Utrecht
Wege H, Müller A, ter Meulen V (1978) Genomic RNA of the murine coronavirus JHM. J Gen Virol 41: 217–227
Wege H, Siddell S, ter Meulen V (1982) The biology and pathogenesis of coronavirus. Curr Top Microbiol Immunol 99: 165–200
Wege H, Koga M, Watanabe R, Nagashima K, ter Meulen V (1983) Neurovirulence of murine coronavirus JHM temperature-sensitive mutants in rats. Infect Immun 39: 1316–1324
Wege H, Dörries R, Wege H (1984) Hybridoma antibodies to the murine coronavirus JHM: Characterization of epitopes on the peplomer protein (E2). J Gen Virol 65: 1931–1942
Weiss SR, Leibowitz JL (1981) Comparison of the RNAs of murine and human coronavirus. In: ter Meulen V, Siddell S, Wege H (eds) Biochemistry and biology and coronaviruses. Plenum, New York, pp 245–259
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Martin, J.P., Koehren, F., Rannou, J.J. et al. Temperature-sensitive mutants of mouse hepatitis virus type 3 (MHV-3): isolation, biochemical and genetic characterization. Archives of Virology 100, 147–160 (1988). https://doi.org/10.1007/BF01487679
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DOI: https://doi.org/10.1007/BF01487679