Interaction of the Retinoblastoma Protein with Orc1 and Its Recruitment to Human Origins of DNA Replication
Figure 3
E2F1 competes with Orc1 for binding to Rb.
(A) Schematic representation of the regions involved in the association of Rb to Orc1 and of those that are necessary for stable association with E2F1. (B) GST pull-down experiment performed by incubating a fixed amount of in vitro translated Orc1 together with scalar amounts of in vitro translated E2F1 with an immobilized GST fusion protein containing the ABC pocket of Rb. The graph shows the quantification of bound E2F1 and Orc1 radioactivity; the input lanes (In) contain the labelled proteins prior to binding. (C) Competitive GST pull-down control experiment performed with in vitro translated Orc1 and luciferase (Luc) proteins using identical experimental conditions as in (B).
