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RNA-dependent RNA Polymerase Activity in Murine Coronavirus-infected Cells

Brian W. J. Mahy

Institut für Virologie und Immunbiologie, Universität Würzburg, Versbacher Strasse 7, D8700 Würzburg, Federal Republic of Germany

The multiplication of murine coronavirus strains A59 or JHM in Sac(-) cells was unaffected by the presence of -amanitin at concentrations which inhibited the host cell DNA-dependent RNA polymerase activity. In cells infected with the A59 virus strain, actinomycin D-resistant RNA synthesis could readily be detected by pulse-labelling with [³H]uridine; this virus-specific RNA synthesis was not induced in the presence of the protein synthesis inhibitor anisomycin. A new RNA-dependent RNA polymerase activity was detected in the large particle fraction of A59 virus-infected cells. Optimal conditions for enzyme activity in vitro were established. Maximum activity occurred 5 h after infection, coincident with the peak of virus-specific RNA synthesis detected by pulse-labelling in vivo.

Keywords: RNA-dependent RNA polymerase, -amanitin, coronavirus replication, Sac(-) cells

Present address: Division of Virology, Department of Pathology, University of Cambridge, Laboratories Block, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2QQ, U.K.

Received 24 May 1982; accepted 29 July 1982.

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