Analysis of the consensus binding sequence and the DNA-binding domain of ZF5

Murine ZF5 is a transcription factor with five zinc finger motifs that represses the c-myc gene by binding to two GC-rich elements at the promoter region. Because of its ubiquitous expression in a variety of tissues, elucidation of biological functions and cellular target genes of ZF5 is of great interest. As the first step of identifying cellular target genes, we have attempted to determine the consensus binding motif for ZF5. We succeeded in isolating 19 oligonucleotide duplex DNAs to which ZF5 binds and determined the binding sequences with DNase I footprinting analysis. From these sequences, we deduced the consensus binding motif for ZF5 to be GSGCGCGR. In addition, we have analyzed the DNA-binding domain of ZF5 by testing a series of deletion mutants. It turned out that the zinc fingers 3 and 4 of the five finger motifs play a critical role in DNA binding.