Figure 2.

Identification of the GZF1 binding consensus sequence. (A) Sequences of 17 cDNA clones recovered in the CASTing analysis. Putative core binding sequences from 17 clones were aligned. (B) EMSA to assess binding of the GST-ΔBTB-GZF1 to selected oligonucleotides. The arrow indicates complexes ofGST-ΔBTB-GZF1 with ³²P-labeled probes (clones 1–14) and the arrowhead indicates free probe. The probes derived from clones 6, 8 and 13 showed very faint or no signals representing the DNA–protein complex. Core sequences of clones 15, 16 and 17 were the same as those of clones 2 (and 4), 10 and 5, respectively (A) and probes from clones 15–17 also showed a strong signal representing the DNA–protein complex (data not shown). (C) Consensus binding sequence of GZF1, as determined by CASTing. The frequency of nucleotides at each position is shown. The putative consensus sequence is 5′-TGCGCN(T/G)(C/A)TATA-3′.