Figure 2

Analyses of the interacting domains. (a) GST pull-down assays using bacterially expressed GST-tagged wild type and various mutants of SUV39H1 and histidine-tagged wild type Tax (His-Tax). A schematic representation of the wild type (SUV39H1-WT) and those of domain structures of mutants are indicated in the upper panels. Results of the pull-down assays are shown in the lower panels. Pulled-down proteins were analyzed by SDS-PAGE and immunoblotting with Lt-4 antibody (top of the lower panels). The bottom panel shows the Coomassie Brilliant Blue (CBB)-stained gel where the wild type and various mutant SUV39H1 proteins were run. (b) Pull-down assays using the wild type GST-SUV39H1 and in vitro translated wild type and various mutant Tax proteins. Schematic description of the structures of wild type and various mutant Tax proteins is presented in the top panel. Results of the pull-down assays are shown in the top of the lower panels. Pulled-down Tax proteins that were labeled with ³⁵S-methionine were visualized by autoradiography (top of the lower panels). The bottom panel shows the autoradiogram of the gel where the radio labeled wild type and various mutant Tax proteins were run.