The operon protects from copper toxicity: CopL is an extracellular membrane-associated copper-binding protein.

As complications associated with antibiotic resistance have intensified, copper (Cu) is attracting attention as an antimicrobial agent. Recent studies have shown that copper surfaces decrease microbial burden, and host macrophages use Cu to increase bacterial killing. Not surprisingly, microbes have evolved mechanisms to tightly control intracellular Cu pools and protect against Cu toxicity. Here, we identified two genes ( and ) encoded within the arginine-catabolic mobile element (ACME) that we hypothesized function in Cu homeostasis. Supporting this hypothesis, mutational inactivation of or increased copper sensitivity. We found that are co-transcribed and that their transcription is increased during copper stress and in a strain in which , encoding a Cu-responsive transcriptional repressor, was mutated. Moreover, displayed genetic synergy with suggesting that CopB functions in Cu export. We further observed that CopL functions independently of CopB or CopA in Cu toxicity protection and that CopL from the clone USA300 is a membrane-bound and surface-exposed lipoprotein that binds up to four Cu ions. Solution NMR structures of the homologous CopL, together with phylogenetic analysis and chemical-shift perturbation experiments, identified conserved residues potentially involved in Cu coordination. The solution NMR structure also revealed a novel Cu-binding architecture. Of note, a CopL variant with defective Cu binding did not protect against Cu toxicity Taken together, these findings indicate that the ACME-encoded CopB and CopL proteins are additional factors utilized by the highly successful USA300 clone to suppress copper toxicity.