The Dissociation Rate of a Winged Helix Protein–DNA Complex Is Influenced by Non-DNA Contact Residues

The dissociation constants and the half-lives of the DNA complexes of winged helix protein Genesis and its mutants were investigated. Our data show that substitutions of non-DNA contact residues in the two highly dynamic wing sequences dramatically change the off rates of the complexes. Thus, our data indicate that the off rate of a DNA complex is encoded in the amino acid sequence of the DNA-binding protein and/or should be a unique property for a protein–DNA complex. Our data also indicate that the local structural transition of a DNA-binding protein in its DNA recognition process is likely to go through a transition state, which can control the on and off rates of a complex.