THE ZNF35 HUMAN ZINC FINGER GENE ENCODES A SEQUENCE-SPECIFIC DNA-BINDING PROTEIN

We developed a rapid method to determine DNA-binding sites for putativ e DNA-binding proteins. This procedure has been successfully used to d efine a specific consensus site for the human ZNF35 zinc finger gene. ZNF35 encodes a 58-kDA polypeptide containing 11 consecutive finger mo tifs located at the amino terminus, and an acidic domain located at th e carboxy terminus. These features suggest that ZNF35 is a site-specif ic DNA-binding protein involved in the regulation of gene expression. We have expressed the ZNF35 protein from E. coli and have employed a S outhwestern-polymerase chain reaction method using random oligonucleot ides to identify its high-affinity binding site. The core sequence for the ZNF35 protein-binding site is 5'-C/GC/GAAG/TA-3'.