title: DNA-binding induces a major structural transition in a type I methyltransferase.
abstract: The type IC DNA methyltransferase M.EcoR124I is a complex multisubunit enzyme that recognizes the non-palindromic DNA sequence GAAN6RTCG. Small angle X-ray scattering has been used to investigate the solution structure of the methyltransferase and of complexes of the enzyme with unmethylated and hemimethylated 30 bp DNA duplexes containing the specific recognition sequence. A major change in the quaternary structure of the enzyme is observed following DNA binding, based on a decrease in the radius of gyration from 56 to 40 A and a reduction in the maximum dimension of the enzyme from 180 to 112 A. The structural transition observed is independent of the methylation state of the DNA. CD shows that there is no change in the secondary structure of the protein subunits when DNA is bound. In contrast, there is a large increase in the CD signal arising from the DNA, suggesting considerable structural distortion which may allow access to the bases targeted for methylation. We propose that DNA binding induces a large rotation of the two HsdM subunits towards the DNA, mediated by hinge bending domains in the specificity subunit HsdS.
authors.affiliation: Biophysics Laboratories, University of Portsmouth, Portsmouth.
authors.forename: I A
authors.initials: IA
authors.lastname: Taylor
authors.forename: K G
authors.initials: KG
authors.lastname: Davis
authors.forename: D
authors.initials: D
authors.lastname: Watts
authors.forename: G G
authors.initials: GG
authors.lastname: Kneale
mesh_terms: D001483:Base Sequence; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D012542:Scattering, Radiation; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000737:chemistry / Q000378:metabolism
chemical_list: D004247:DNA; C072994:EcoR124 modification methylase type I; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)