title: Substitution of the conserved phenylalanine in the S-adenosyl-L-methionine binding site of M.MspI with tyrosine modifies the kinetic properties of the enzyme.
abstract: Cytosine (C-5)-specific DNA methyltransferases share a set of ten conserved motifs distributed evenly throughout the entire polypeptide chain. The first conserved motif contains a Phe, which is intimately associated with cofactor recognition. In the pseudo-DNA methyltransferase M.SpoI, encoded by the pmt1 gene in Schizosaccharomyces pombe, a Tyr replaces this Phe residue. We describe the properties of a mutant form of M.MspI, a typical cytosine (C-5)-specific DNA methyltransferase, in which Tyr replaces the conserved Phe. This mutant shows differences in ternary complex formation and in the pattern of covalent complex formation with an inhibitory, fluorinated DNA duplex which may be due to anomalous hydrogen bonding between the mutant Tyr hydroxyl group and the catalytic loop of the enzyme or through interference with cofactor binding.
authors.affiliation: Krebs Institute for Biomolecular Research, Department of Molecular Biology, University of Sheffield, Western Bank, UK.
authors.forename: E
authors.initials: E
authors.lastname: Pinarbasi
authors.forename: M S
authors.initials: MS
authors.lastname: Kan
authors.forename: C
authors.initials: C
authors.lastname: Duran
authors.forename: G C
authors.initials: GC
authors.lastname: Ford
authors.forename: D P
authors.initials: DP
authors.lastname: Hornby
mesh_terms: D001665:Binding Sites; D015257:DNA-Cytosine Methylases / Q000235:genetics* / Q000378:metabolism*; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine / Q000235:genetics / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000378:metabolism*; D012568:Schizosaccharomyces; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*
chemical_list: D014443:Tyrosine; D010649:Phenylalanine; D012436:S-Adenosylmethionine; C029641:DNA modification methylase HpaII; D015257:DNA-Cytosine Methylases