Figure 1. Interaction between β-catenin and CBP. A, Complementation of the cdc25-2 mutation through the interaction of β-catenin COOH-terminal region with CBP. The temperature-sensitive cdc25-2 yeast cells were cotransformed with the indicated plasmids and the CBP expression plasmid isolated in the screening, and incubated on galactose containing plates either at 25 (left) or 37°C (right). B, Fine mapping of the β-catenin domain that binds to CBP in yeast. Cells were cotransformed with the indicated plasmids together with the CBP expression plasmid, and tested for the growth at 37°C as described for A. The numbers indicate the locations in the amino acid sequence of β-catenin. ++, Efficient growth; +, poor growth; −, no growth. C, β-Catenin directly binds to CREB-binding domain of CBP. Bacterially expressed and purified His-tagged βcatR10-C or myc-tagged β-catenin was incubated with buffer (lane 5), GST (lanes 2 and 6), or GST-CBP 451–682 (lanes 3 and 7). After extensive washing, bound proteins were eluted and resolved by SDS-PAGE. βCatR10-C and β-catenin were detected by an anti-His antibody and an anti-myc antibody, respectively. Approximately 10% of the input is shown (lanes 1 and 4). D, Coimmunoprecipitation of CBP and β-catenin. COS7 cells were transfected with expression plasmids for CBP and/or myc-tagged pt β-catenin as indicated. Cell lysates were immunoprecipitated with anti-CBP polyclonal antibodies 2 d after transfection. The immunoprecipitates were separated by SDS-PAGE and subjected to Western blotting with anti-myc antibodies.
Image published in: Takemaru KI and Moon RT (2000)
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