Mapping the 5' ends, 3' ends, and splice sites of mRNAs from the early E3 transcription unit of adenovirus 5

Abstract

Using nuclease gel analyses, the sites in the approximately 4000 nucleotide (nt) E3 transcription unit of adenovirus 5 (Ad5) that encode the 5' ends, 3' ends, and 5' and 3' splice sites of the approximately 10 E3 mRNAs were determined. Transcription initiation of all mRNAs occurs at two major (nt 1 and 8) and approximately two minor sites, situated 20-30 nt 3' to a TATA box. There are two major 3' end sites (nt 2227 and 3308), located approximately 20 nt downstream from ATTAAA and an AATAAA sequences, respectively. Thus, ATTAAA, as well as the usual AATAAA, apparently can function as a 3' end signal. There are two 5' splice sites (nt 372 and 923), both with GT at the intron boundary. There are four 3' splice sites (nt 766, 1817, 2201, and 2880), all with AG at the intron boundary. The nt 1817, 2201, and 2880 3' splice sites are located immediately upstream from open reading frames, such that splicing at the different sites allows synthesis of completely different proteins.