DNA-binding activities of GST-fused DNA-binding domain 1 and 2 of MEL1 against DNA-binding consensus sequences. DNA-binding activities of each GST fusion protein were detected by electrophoretic mobility shift assay (EMSA). End-labeled consensus sequences of DNA-binding domain 1 (D1-CONS) and DNA-binding domain 2 (D2-CONS) were used for EMSA; 100 ng of each purified GST fusion protein was incubated with ³²P-labeled D1- or D2-CONS oligonucleotides. ³²P-labeled D1-CONS was incubated with control GST (lane 1), EVI1DBD1 (lane 2), or MEL1DBD1 (lane 5) and ³²P-labeled D2-CONS was incubated with control GST (lane 8), EVI1DBD2 (lane 9), or MEL1DBD2 (lane 12). For the competition assay, a 100-times higher concentration of double-stranded D1-CONS (D1; lanes 3 and 6) or D2-CONS (D2; lanes 10 and 13) or else the same amount of double-stranded random oligonucleotides (NSP; lanes 4, 7, 11, and 14) was incubated in each reaction.