FIG. 3.

Mapping of the minimal domain of E1 that can be recruited to the origin by E2. (A) Diagram of the wild-type E1 protein indicating the positions of conserved regions A, B, C, and D and of the DNA-binding domain. Truncated mutant E1 proteins are diagrammed, and the amino acids at the N and C terminus of each truncated protein are indicated. (B and C) E2-dependent origin-binding assay with truncated E1 proteins. DNA-protein complexes were assembled with E2 and the indicated E1 protein made by in vitro translation. The immunoprecipitation of the complexes was performed with the rabbit K72 polyclonal antibody raised against the C-terminal 15 amino acids of HPV-11 E1 (B) or with the M2 anti-FLAG monoclonal antibody (C). The proteins shown in panel C were tagged at their N termini with the FLAG epitope. Detection of bound DNA was done as described in the legend to Fig. 2A. −E1, no E1 present.