Cis- and trans-acting factors for transcription of the adenovirus 12 E1A gene

Cis- and trans-acting factors were analyzed for transcription of the adenovirus 12 E1A gene possessing two sites for transcription initiation. These sites are located at nucleotide positions 306 and 445 with respect to the left end of the viral genome as position 1. The template activity of DNAs with various deletions at the 5'-upstream region of the E1A gene was examined in a cell-free system using a nuclear extract of Ehrlich ascites tumor cells. A DNA region specifically stimulating transcription initiated at the site distal to the E1A coding sequence was found located between positions 1 and 166. No DNA sequence affecting transcription from a proximal start-site appeared to be present in the region between positions 1 and 378. DNaseI-footprinting indicated that factors present in the extract bind to two distinct DNA segments, both of which are located within a region stimulating distal transcription. Two footprints were observed, one between positions 19 and 55 and the other between 77 and 94. The former footprint was inhibited by synthetic oligonucleotides containing a sequence recognized by nuclear factor I and the latter contained a sequence similar to one present in the B-enhancer of polyoma virus. Competition of in vitro transcription with synthetic oligonucleotides indicated (a) nuclear factor(s) bound to the region between positions 19 and 55 to be responsible for stimulating distal transcription of the adenovirus 12 E1A gene.