Figure 4.

In vitro binding of the recombinant BPBF protein to the pyrimidine box sequence motif. EMSA performed with affinity purified GST-BPBF protein and oligonucleotides deduced from the thiol protease AL21 (A; D₁ motif in Fig. 5) and the α-amylase Amy2/32b gene promoters (B). The sequences of the corresponding wild-type (WT) and mutant derivative (mt) oligonucleotides, used as probes, are shown at the bottom of each panel. The pyrimidine box motif is underlined and the point mutations are indicated by lowercase letters. Binding reactions were performed with the GST protein as a negative control (C) or with the GST-BPBF protein (+), in the absence (−) or in the presence of probe competitors at the indicated molar excesses (20×, 50×, and 100×).