Mechanism and requirements for bovine papillomavirus, type 1, E1 initiator complex assembly promoted by the E2 transcription factor bound to distal sites.

DNA replication of papillomavirus requires the viral initiator E1 and the transcription factor E2. Bovine papillomavirus, type 1 (BPV-1), E1, and E2 bind cooperatively as dimers to proximal sites in the viral replicator generating a sequence-specific E1E2-ori complex. This complex is critical for replication and can be converted to a multimeric E1-ori initiator complex by displacement of E2 in the presence of hydrolyzable ATP. However, E2 can function over extended distances, and E2 at a distal position 33 base pairs upstream of the E1-binding site also loads an E1 dimer onto ori. Under these conditions, neither displacement of E2 nor ATP hydrolysis are required for E1-ori formation, consistent with a need for ATP hydrolysis in E2 displacement from E1E2-ori. However, ATP is required for stabilization of the resulting E1-ori complex. These results indicate that BPV (with a proximal E2-binding site) and human papillomaviruses (with distal E2-binding sites) utilize the same general mechanism for E1 loading but suggest that E1E2-ori, which forms preferentially on ori, may perform an additional role in BPV replication.