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Infection of terminally differentiated myotubes with Rous sarcoma virus (RSV): lack of DNA integration but presence of RSV mRNA

Akio Tanaka

Jan Wolfert^>

Mitsuhiro Fujihara

Department of Microbiology, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, U.S.A.

We re-examined the generally accepted concept that replication of Rous sarcoma virus (RSV) requires host DNA synthesis. We used terminally differentiated chicken myotubes as the host because chromosomal DNA replication is completely abolished by the natural differentiation process. Southern blot analysis detected unintegrated viral DNA in both the nucleus and cytoplasm of infected myotubes. This indicated that reverse transcription of the infecting viral RNA and transport of the newly synthesized viral DNA into the nucleus proceeded normally in myotubes. However, restriction enzyme digestion of high M_r DNA prepared from infected myotubes produced none of the fragments specific for RSV, indicating that the viral DNA had failed to integrate into the myotube chromosomal DNA. In these infected myotubes, viral RNA was detected by in situ hybridization. Northern blot analysis showed the presence of all three RSV mRNAs (38S, 28S and 21S). The amount of these viral RNAs in infected myotubes was comparable with that found in infected fibroblasts. We conclude that host DNA synthesis is required for RSV integration, but, in contrast to the generally accepted concept, viral DNA integration is not an absolute requirement for transcription of the RSV genome.

Present address: Center for Molecular Medicine and Immunology, 1 Bruce Street, Newark, New Jersey 07103, U.S.A.

^> Present address: Baxter Pandex Division, 909 Orchard Street, Mundelein, Illinois 60060, U.S.A.

Present address: Department of Microbiology, Molecular Genetics and Immunology, School of Medicine, University of Kansas Medical Center, Kansas City, Kansas 66103, U.S.A.

Received 16 May 1991; accepted 14 February 1992.

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