Figure 1.

HPV E1 physically interacts with DNA polymerases ϵ and δ. (A) Enzyme-linked Immunoassays (ELISAs) were performed using 100 ng of pol ϵ, pol δ or BSA as the immobilized protein. After washing, increasing amounts of MBP-E1 (as indicated) were added to the wells. Binding was detected using a polyclonal anti-MBP antibody followed by a secondary HRP-conjugated antibody. After washing, and addition of HRP substrate, absorbance was measured at 450 nm. Error bars are defined as Standard Deviation (s.d.); n = 6. (B) Far western blot analysis was performed by subjecting 10 μg of single-step affinity purified pol ϵ or pol δ to electrophoresis, transferring and then probing the membrane with 10 μg/ml of either GST-E1 (first panel) or GST (second panel) (100 μg total), according to the Materials and Methods section. A monoclonal anti-GST antibody was used as the primary antibody, and detected using HRP-linked secondary antibody as described in the Materials and Methods.