CopZ from Bacillus subtilis interacts in vivo with a copper exporting CPx-type ATPase CopA
David S. Radford
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
Search for more papers by this authorMargaret A. Kihlken
School of Chemical Sciences and Pharmacy, University of East Anglia, Norwich NR4 7TJ, UK
Search for more papers by this authorGilles P.M. Borrelly
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
Search for more papers by this authorColin R. Harwood
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
Search for more papers by this authorNick E. Le Brun
School of Chemical Sciences and Pharmacy, University of East Anglia, Norwich NR4 7TJ, UK
Search for more papers by this authorCorresponding Author
Jennifer S. Cavet
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
*Corresponding author. Tel.: +44 (191) 222 5809; Fax: +44 (191) 222 7736, E-mail address: [email protected]Search for more papers by this authorDavid S. Radford
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
Search for more papers by this authorMargaret A. Kihlken
School of Chemical Sciences and Pharmacy, University of East Anglia, Norwich NR4 7TJ, UK
Search for more papers by this authorGilles P.M. Borrelly
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
Search for more papers by this authorColin R. Harwood
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
Search for more papers by this authorNick E. Le Brun
School of Chemical Sciences and Pharmacy, University of East Anglia, Norwich NR4 7TJ, UK
Search for more papers by this authorCorresponding Author
Jennifer S. Cavet
School of Cell and Molecular Biosciences, The Medical School, University of Newcastle, Newcastle NE2 4HH, UK
*Corresponding author. Tel.: +44 (191) 222 5809; Fax: +44 (191) 222 7736, E-mail address: [email protected]Search for more papers by this authorAbstract
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. ΔcopA 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 caa3 oxidase was retained in ΔcopZ and ΔcopA. ΔcopZ was only slightly copper-hypersensitive but ΔcopZ/ΔcopA was more sensitive than ΔcopA, implying some action of CopZ that is independent of CopA. Significantly, ΔcopZ 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.
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