High-level transcription from the adenovirus major late promoter requires downstream binding sites for late-phase-specific factors.

The adenovirus major late promoter (MLP) is active during both the early and late phases of infection. During the early phase the activity of the MLP is similar to those of the other early viral promoters, but during the late phase the rate of transcription from the MLP becomes much greater by comparison. We report here that sequence-specific binding proteins are induced during the late phase which interact with three regions in the first intron of the MLP transcription unit from positions +37 to +68, +80 to +105, and +105 to +125 relative to the transcription initiation site. To measure the significance of these binding sites for transcription during the late phase, we constructed MLP-beta-globin fusions and substituted them for early region 3 in adenovirus recombinants. Deletion of the binding sites caused significant reductions in the rate of transcription, specifically during the late phase of infection. Deletion of all three sites reduced the rate of transcription 25- to 50-fold and the accumulation of cytoplasmic MLP-beta-globin RNA 200-fold. These results indicate that the high rate of transcription from the MLP during the late phase of infection results from the interaction of virus-induced transcription factors with three binding sites in the first intron of the major late transcription unit.