Organization Of Rna Transcripts From A 7.8-Kb Region Of The Frog Virus-3 Genome

The detailed organization of the RNAs transcribed from a region of the FV 3 genome ( Sa /l-F fragment and adjacent sequences) has been determined. The information was derived from the cell-free translation of hybrid-selected RNA to locate the genes encoding specific polypeptides, RNA filter hybridization to size the transcripts, and S1 nuclease mapping to locate the 5′- and 3′-ends of the RNAs on the genome. Three genes are contiguous and are transcribed from the same strand: two immediate early genes encoding transcripts of about 1.3 kb that directed the in vitro synthesis of 42K and 46K polypeptides, separated by the late gene encoding the major capsid protein (48K). At an advanced stage in infection, transcripts derived from the immediate early genes are also present. A set of RNAs with different 5′-ends ranging from 1.7 to 0.58 kb is produced from the p46 gene region whereas RNAs, 0.98 and 0.6 kb in size, complementary to the 5′-end of the p42 message, are synthesized. This gene cluster is located between two genes transcribed in the opposite direction from the rightward-reading strand: a late gene whose message is 0.5 kb in size and encodes a 15K polypeptide and a gene transcribed at immediate early and late times of infection which encodes a protein of 70 kDa. The 5′-end of the late RNA maps downstream of the 5′-end of the early one, their sizes being 1.85 and 2 kb, respectively, but both of them can be translated in vitro into a 70K polypeptide. These observations suggest that transcription is not regulated by the organization of the genes; they suggest rather that specific DNA sequences are responsible for the promotion of immediate early and late transcriptions.