Sequence-Selective and Hydrolytic Cleavage of DNA by Zinc Finger Mutants

摘要: We have reported the successful conversion of structural zinc site in finger peptides to a functional site. A series resulting mutants exhibit hydrolytic ability activated ester depending on coordination geometry and acidity ions. In this study, we explored DNA by H4 mutant since showed highest among peptides. The zinc-bound form peptide exhibited phosphoesters even converted supercoiled plasmid nicked circular form. An increasing ionic strength leads loss nuclease due nonspecific interaction between DNA. sharp contrast, three-tandem H4-type protein performed specific hydrolysis at GC box high strength. Thus, present study demonstrated that converting native is novel approach for creating artificial nucleases with sequence selectivity.