The linker region, the carboxyl-terminal zinc finger, and the basic arm of GATA-1 are sufficient for binding p53TAD in an in vitro pulldown assay. (A) Schematic of GATA-1 highlighting the zinc-finger domains (GATA-1 ZFD) and the various deletion mutants of GATA-1 ZFD used as GST fusion proteins. (B) Comparative binding of GST-GATA-1 ZFD and GST-GATA-1 NF plus L (GST-GATA-1_200-251) to p53TAD. Various concentrations (1.0, 0.1, and 0.01 μM) of p53TAD were incubated with either 1 μM GST-GATA-1 ZFD (lanes 3-5) or GST-GATA-1 NF plus L (lanes 6-8). In the GST lane, 1 μM purified p53TAD was incubated with 1 μM GST as a control (lane 2). The input lane is 0.5% p53TAD (lane 1). (C) Comparative binding of GST-GATA-1 ZFD and GST-GATA-1 L plus CF (GST-GATA-1_228-317) to p53TAD. Various concentrations (1.0, 0.1, and 0.01 μM) of p53TAD were incubated with either 1 μM GST-GATA-1 ZFD (lanes 3-5) or GATA-1 L plus CF (lanes 6-8). In the GST lane, 1 μM purified p53TAD was incubated with 1 μM GST as a control (lane 2). The input lane is 0.5% p53TAD (lane 1). (D) Comparative binding of GST-GATA-1 ZFD and GST-GATA-1 CF (GST-GATA-1_252-317) to p53TAD. Various concentrations (1.0, 0.1, and 0.01 μM) of p53TAD were incubated with either 1 μM GST-GATA-1 ZFD (lanes 3-5) or GATA-1 CF plus BA (lanes 6-8). In the GST lane, 1 μM purified p53TAD was incubated with 1 μM GST as a control (lane 2). The input lane is 0.5% p53TAD (lane 1). In all experiments, bound protein was detected with an anti-p53 antibody DO-1.