title: CopZ from Bacillus subtilis interacts in vivo with a copper exporting CPx-type ATPase CopA.
abstract: The structure of the hypothetical copper-metallochaperone CopZ from Bacillus subtilis and its predicted partner CopA have been studied but their respective contributions to copper export, -import, -sequestration and -supply are unknown. DeltacopA was hypersensitive to copper and contained more copper atoms cell(-1) than wild-type. Expression from the copA operator-promoter increased in elevated copper (not other metals), consistent with a role in copper export. A bacterial two-hybrid assay revealed in vivo interaction between CopZ and the N-terminal domain of CopA but not that of a related transporter, YvgW, involved in cadmium-resistance. Activity of copper-requiring cytochrome caa(3) oxidase was retained in deltacopZ and deltacopA. DeltacopZ was only slightly copper-hypersensitive but deltacopZ/deltacopA was more sensitive than deltacopA, implying some action of CopZ that is independent of CopA. Significantly, deltacopZ contained fewer copper atoms cell(-1) than wild-type under these conditions. CopZ makes a net contribution to copper sequestration and/or recycling exceeding any donation to CopA for export.
authors.affiliation: School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK.
authors.forename: David S
authors.initials: DS
authors.lastname: Radford
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Kihlken
authors.forename: Gilles P M
authors.initials: GP
authors.lastname: Borrelly
authors.forename: Colin R
authors.initials: CR
authors.lastname: Harwood
authors.forename: Nick E
authors.initials: NE
authors.lastname: Le Brun
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Cavet
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism*; D020816:Amino Acid Motifs; D001412:Bacillus subtilis / Q000187:drug effects / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D003300:Copper / Q000378:metabolism* / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017136:Ion Transport; D008954:Models, Biological; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D016296:Mutagenesis; D025941:Protein Interaction Mapping; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: D001426:Bacterial Proteins; D027682:Cation Transport Proteins; C071263:CopA protein, Bacteria; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D003300:Copper; D000251:Adenosine Triphosphatases