Transcription and processing of adenoviral RNA by extracts from HeLa cells

Concentrated extracts from HeLa cells prepared by the procedure of Sugden and Keller [Sugden, B. & Keller, W. (1973) J. Biol. Chem. 248, 3777-3788] can synthesize RNA molecules of discrete lengths in the presence of DNA templates carrying complete viral transcription units. Accurate initiation of transcription was shown to occur at the "major late" promoter and at the promoter for the "early region III" transcription unit of the adenovirus 2 genome. A cloned fragment of adenovirus 2 DNA containing the early region III transcription unit directs the synthesis of discrete RNA species, many of which correspond in length to spliced early region III mRNAs isolated from adenovirus 2-infected HeLa cells. As shown by hybridization/nuclease S2 analysis, RNA splicing and the formation of specific 3' ends are also taking place in vitro.