SEQUENCE-SPECIFIC RECOGNITION BY CYTOSINE C-5 AND ADENINE N-6 DNA METHYLTRANSFERASES REQUIRES DIFFERENT DEFORMATIONS OF DNA

DNA methyltransferases modify specific cytosines and adenines within 2 -6 bp recognition sequences. We used scanning force microscopy and gel shift analysis to show that M.HhaI, a cytosine C-5 DNA methyltransfer ase, causes only a 2 degrees bend upon binding its recognition site, O ur results are consistent with prior crystallographic analysis showing that the enzyme stabilizes an extrahelical base while leaving the DNA duplex otherwise unperturbed. In contrast, similar analysis of M.EcoR I, an adenine N-6 DNA methyltransferase, shows an average bend angle o f approximately 52 degrees. This distortion of DNA conformation by M.E coRI is shown to be important for sequence-specific binding.