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 repr esses the c-myc gene by binding to two GC-rich elements at the promoter reg ion. Because of its ubiquitous expression in a variety of tissues, elucidat ion of biological functions and cellular target genes of ZF5 is of great in terest. As the first step of identifying cellular target genes, we have att empted to determine the consensus binding motif for ZF5. We succeeded in is olating 19 oligonucleotide duplex DNAs to which ZF5 binds and determined th e binding sequences with DNase I footprinting analysis. From these sequence s, we deduced the consensus binding motif for ZF5 to be GSGCGCGR. In additi on, we have analyzed the DNA-binding domain of ZF5 by testing a series of d eletion mutants. It turned out that the zinc fingers 3 and 4 of the five fi nger motifs play a critical role in DNA binding. (C) 1999 Academic Press.