title: Surface labelling of the type I methyltransferase M.EcoR124I reveals lysine residues critical for DNA binding.
abstract: The type IC methyltransferase M.EcoR124I consists of a specificity subunit (HsdS) and two methylation subunits (HsdM). Using chemical modifications, we have investigated the accessibility of lysine residues in the free enzyme and in the complex with its DNA recognition sequence. A total of 41 of the 109 lysine residues in the enzyme are susceptible to modification, of which 19 are located in the HsdS subunit and 11 in each of the two HsdM subunits. DNA binding results in extensive protection of lysine residues in the HsdS subunit, while those in the HsdM subunit are only protected weakly. The DNA binding activity of the methylase is abolished when a small fraction of the accessible lysine residues are modified. Peptide mapping and N-terminal sequencing has been used to locate the rapidly modified lysine residues in HsdS that are critical for DNA binding. Highly modified residues (K297, K261 and K327) are found in the C-terminal variable domain that is responsible for DNA recognition, but others (K196, K203 and K210) are found in the conserved regions that had not previously been implicated in DNA binding.
authors.affiliation: Biophysics Laboratories, School of Biological Sciences, University of Portsmouth, UK.
authors.forename: I A
authors.initials: IA
authors.lastname: Taylor
authors.forename: M
authors.initials: M
authors.lastname: Webb
authors.forename: G G
authors.initials: GG
authors.lastname: Kneale
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism*; D008239:Lysine / Q000032:analysis / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D017421:Sequence Analysis; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000737:chemistry / Q000378:metabolism; D000521:alpha-MSH / Q000378:metabolism
chemical_list: D000596:Amino Acids; D010446:Peptide Fragments; D000521:alpha-MSH; D004247:DNA; C072994:EcoR124 modification methylase type I; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D008239:Lysine