CopZ from Bacillus subtilis interacts in vivo with a copper exporting CPx-type ATPase CopA.
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.
Radford
David S
DS
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK.
Kihlken
Margaret A
MA
Borrelly
Gilles P M
GP
Harwood
Colin R
CR
Le Brun
Nick E
NE
Cavet
Jennifer S
JS
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
D001426:Bacterial Proteins; D027682:Cation Transport Proteins; C071263:CopA protein, Bacteria; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D003300:Copper; D000251:Adenosine Triphosphatases