title: Zinc binding by the methylation signaling domain of the Escherichia coli Ada protein.
abstract: The Escherichia coli Ada protein repairs O6-methylguanine residues and methyl phosphotriesters in DNA by direct transfer of the methyl group to a cysteine residue located in its C- or N-terminal domain, respectively. Methyl transfer to the N-terminal domain causes it to acquire a sequence-specific DNA binding activity, which directs binding to the regulatory region of several methylation-resistance genes. In this paper we show that the N-terminal domain of Ada contains a high-affinity binding site for a single zinc atom, whereas the C-terminal domain is free of zinc. The metal-binding domain is apparently located within the first 92 amino acids of Ada, which contains four conserved cysteine residues. We propose that these four cysteines serve as the zinc ligand residues, coordinating the metal in a tetrahedral arrangement. One of the putative ligand residues, namely, Cys69, also serves as the acceptor site for a phosphotriester-derived methyl group. This raises the possibility that methylation-dependent ligand reorganization about the metal plays a role in the conformational switching mechanism that converts Ada from a non-sequence-specific to a sequence-specific DNA-binding protein.
authors.affiliation: Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138.
authors.forename: L C
authors.initials: LC
authors.lastname: Myers
authors.forename: M P
authors.initials: MP
authors.lastname: Terranova
authors.forename: H M
authors.initials: HM
authors.lastname: Nash
authors.forename: M A
authors.initials: MA
authors.lastname: Markus
authors.forename: G L
authors.initials: GL
authors.lastname: Verdine
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry*; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism; D029968:Escherichia coli Proteins; D008745:Methylation; D008969:Molecular Sequence Data; D019853:O(6)-Methylguanine-DNA Methyltransferase; D013329:Structure-Activity Relationship; D014157:Transcription Factors; D015032:Zinc / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D014157:Transcription Factors; C017800:zinc-binding protein; C045675:Ada protein, E coli; D019853:O(6)-Methylguanine-DNA Methyltransferase; D015032:Zinc