CusF was identified in only five families and in 62% of them it co-localized with cusABC. However, the fact that in 22 organisms CusB and CusF were fused in a single gene do not compare with the role of CusF as a soluble carrier, a role that certainly deserves to be revised. In E. coli APEC 01 we identified a CusABC paralog, named SilABC which is plasmid borne and adjacent to PcoAB, with an apparent role in silver extrusion suggesting evolution by duplication and functional equivalence but metal-binding specialization. These analyses were performed with the aim to elucidate between Pevonedistat cell line two hypotheses for the concurrent evolution of well characterized

interacting protein sets in copper homeostasis: function dominance or protein-protein interaction dominance, The high presence correlation of CusABC support protein-protein interaction as the selection trait for the assembly with two caveats: CusC may still be functional in the absence TGF-beta cancer of CusAB (as happens in other RND groups, [43]). This idea is consistent with the fact that in a number of cases cusC was found to lie adjacent to genes encoding for RND complexes with other proposed specificities. Additionally it would be interesting to determine if the minimal set of an inner Captisol chemical structure membrane protein such CopA and a single outer membrane protein such as CusC

are sufficient for copper tolerance Sodium butyrate acquisition. In contrast, the low presence correlation between

PcoA/PcoC compared to the higher and unexpected correlation of PcoC with CueO may lead to observation that CueO functionally replaces PcoA on the interaction with PcoC. However, CueO and PcoA belong to the MCO structural family and, in spite of sharing low identity at the sequence level, their three dimensional structure is highly preserved as happens with the rest of the family members [44]. In both cases evidence support the protein-protein interaction hypothesis as the basic mechanisms for the evolution of the copper homeostasis systems supporting our theoretical treatment as metabolic networks [45]. Conclusions Our results suggest complex evolutionary dynamics and still unexplored interactions among different proteins to achieve copper homeostasis in gamma proteobacteria, challenging some of the molecular transport mechanism proposed for these systems. Methods Gamma proteobacterial genomes To carry out this analysis we analyzed 268 proteobacterial genomes available from the KEGG database (Release 56.0, October 1, 2010) [46, 47] (Aditional file 1). Protein sequences used as seeds for ortholog detection CopA from Escherichia coli K-12 MG1655 [KEGG:eco:b0484]; CueO from Escherichia coli O1:K1:H7 (APEC) [KEGG:ecv:APECO1_1862]; CueP from Salmonella enterica subsp.