A new structural paradigm in copper resistance in Streptococcus pneumoniae.
Copper resistance has emerged as an important virulence determinant of microbial pathogens. In Streptococcus pneumoniae, copper resistance is mediated by the copper-responsive repressor CopY, CupA and the copper-effluxing P(1B)-type ATPase CopA. We show here that CupA is a previously uncharacterized cell membrane-anchored Cu(I) chaperone and that a Cu(I) binding-competent, membrane-localized CupA is obligatory for copper resistance. The crystal structures of the soluble domain of CupA and the N-terminal metal-binding domain (MBD) of CopA (CopA(MBD)) reveal isostructural cupredoxin-like folds that each harbor a binuclear Cu(I) cluster unprecedented in bacterial copper trafficking. NMR studies reveal unidirectional Cu(I) transfer from the low-affinity site on the soluble domain of CupA to the high-affinity site of CopA(MBD). However, copper binding by CopA(MBD) is not essential for cellular copper resistance, consistent with a primary role of CupA in cytoplasmic Cu(I) sequestration and/or direct delivery to the transmembrane site of CopA for cellular efflux.
Department of Chemistry, Indiana University, Bloomington, Indiana, USA.
Yue
Y
Fu
Ho-Ching Tiffany
HC
Tsui
Kevin E
KE
Bruce
Lok-To
LT
Sham
Khadine A
KA
Higgins
John P
JP
Lisher
Krystyna M
KM
Kazmierczak
Michael J
MJ
Maroney
Charles E
CE
Dann
Malcolm E
ME
Winkler
David P
DP
Giedroc
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003300:Copper / Q000378:metabolism / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary; D013296:Streptococcus pneumoniae / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism / Q000472:pathogenicity
D001426:Bacterial Proteins; C071263:CopA protein, Bacteria; D003300:Copper