FIGURE 1.

Interaction between MCM7 and cyclin E/Cdk2. A, 293T cells were transfected with FLAG-MCM7 together with myc-cyclin E and myc-Cdk2. Cell lysates were immunoprecipitated (IP) with myc beads followed by immunoblotting (IB) with FLAG antibody. B, 293T cells were transfected with the indicated plasmids, and the cell lysates were immunoprecipitated with FLAG beads followed by immunoblotting with myc antibody. C, 293T cells were transfected with myc-cyclin E and myc-Cdk2. The cell lysates were subjected to a pull-down assay with GST or GST-MCM7 and immunoblotted with myc antibody. D, 293T cell lysates were immunoprecipitated with Cdk2 antibody or normal mouse IgG and then immunoblotted with MCM7 and cyclin E antibodies. E, GST or GST-MCM7 proteins immobilized on glutathione beads were incubated with His-Cdk2 in lysis buffer for 2 h. The associated protein was eluted with SDS loading buffer and immunoblotted with Cdk2 antibody. F, MCM7 and its RXL mutants were generated as indicated and transfected into 293T cells together with myc-cyclin E/Cdk2. Cell lysates were immunoprecipitated with FLAG beads and eluted for immunoblotting with myc antibody. TCE, total cell extract; NMS, normal mouse serum.