title: Identification of the primary metal ion-activation sites of the diphtheria tox repressor by X-ray crystallography and site-directed mutational analysis.
abstract: The diphtheria tox repressor, DtxR, is a 226 amino acid transition metal ion-activated regulatory protein that controls the expression of diphtheria toxin in toxigenic Corynebacterium diphtheriae. The previously solved three-dimensional DtxR structures have identified two potential metal ion binding sites which may play a role in the activation of DNA binding by the repressor. We have used both X-ray crystallographic and site-directed mutational analysis of DtxR(C102D)-Ni2+ complexes and DtxR to identify the metal ion-binding site which results in the activation of the repressor. We demonstrate that DtxR contains both a primary and an ancillary metal ion binding site. The primary site functions directly in the activation of DNA binding. In contrast, the ancillary site contributes weakly, if at all, to activation.
authors.affiliation: Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02154, USA.
authors.forename: X
authors.initials: X
authors.lastname: Ding
authors.forename: H
authors.initials: H
authors.lastname: Zeng
authors.forename: N
authors.initials: N
authors.lastname: Schiering
authors.forename: D
authors.initials: D
authors.lastname: Ringe
authors.forename: J R
authors.initials: JR
authors.lastname: Murphy
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002413:Cations, Divalent; D003353:Corynebacterium diphtheriae / Q000737:chemistry*; D018360:Crystallography, X-Ray; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009532:Nickel / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D001426:Bacterial Proteins; D002413:Cations, Divalent; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C075586:DtxR protein, Corynebacterium diphtheriae; D009532:Nickel