pestis transcriptional profiling studies where increased bfr expression and, in one case, decreased ftnA expression were reported for iron-limiting growth environments [33, 35]. Post-transcriptional regulatory functions in iron-deficient cells have also been attributed to aconitases. In fact, NVP-BSK805 chemical structure eukaryotic AcnA has been termed iron-responsive protein 1 (IRP-1) [60]. Apo-enzyme versions of E. coli aconitases stabilize their cognate mRNAs

and influence the expression of sodA. AcnA enhanced sodA transcript stability and was induced by iron starvation and find protocol oxidative stress in E. coli [61, 62]. These findings could not be easily reconciled with our data onAcnA and AcnB abundance changes in Y. pestis. AcnA and AcnB were decreased in abundance, as were the combined aconitase activities, in iron-depleted cells. SodA abundance was not significantly affected by either growth phase [39] or iron depletion. The response

of Y. pestis to iron starvation and cellular stress resulting from the loss of this metal ion seems to implicate a network of regulators, as presented in Figure 5. Indeed, functional relationships between Fur and OxyR [32], Fur and CRP [31] and Fur and p38 MAPK pathway apo-aconitases [62] were previously reported for E. coli. Iron starvation stress responses Numerous E. coli genes encoding oxidative stress response proteins are co-regulated by SoxR, Fur and OxyR according to information in the ZD1839 EcoCyc database. The OxyR H2O2-response system restored Fur repression in iron-replete media during oxidative stress in E. coli [32], a mechanism that we think is also relevant in Y. pestis. Strong abundance decreases in iron-starved Y. pestis cells were observed for three iron-dependent proteins, SodB, KatE and KatY. The three enzymes detoxify peroxides and radicals formed during oxidative stress. Proteins with similar functions but cofactors other than

iron (e.g. SodA and AhpC) were not markedly changed in abundance. Functional assays supported such proteomic data; SOD activities in iron-depleted cells dropped markedly less than catalase activities. In conclusion, our data strongly support the notion that Y. pestis adapts its repertoire of oxidative stress response enzymes by limiting the expression of iron cofactor-dependent enzymes, when iron is in short supply. The coordination of bacterial responses to iron limitation and the defence against oxidative stress was proposed earlier [63]. Iron acquisition systems All Y. pestis biovars have several proven iron acquisition systems, and transcriptional control by Fur has been demonstrated [18, 64]. The genes and operons for putative iron transporters (e.g. Ysu, Fit, Fhu, Iuc, Has) also feature conserved 19-nt Fur-binding sites to which recombinant Fur binds [20].