E2A Antagonizes PU.1 Activity through Inhibition of DNA Binding

Figure 2

Identification of domains required for mediating the interaction between PU.1 and E47. (a) In vitro translated ³⁵S-methionine labeled full-length human E47 (FL, aa 1–654), E47 C-terminal truncation 1 (CT1, aa 1–492), and E47 C-terminal truncation 2 (CT2, aa 1–369) were incubated with bacterially produced GST-PU.1 fusion protein. 10% input proteins and proteins bound to GST-PU.1 were separated by SDS-PAGE and visualized by autoradiography. (b) ³⁵S-methionine labeled in vitro translated PU.1 and E47 were incubated with GST-E47bHLH protein (E47, aa 521–623). In vitro translated E47 was used as a positive control since it should dimerize with the E47 bHLH domain. (c) ³⁵S-methionine labeled in vitro translated PU.1 deletion mutants were incubated with GST-E47bHLH protein.

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