Role of a TonB-dependent receptor and an oxygenase in iron-dependent copper resistance in <i>Caulobacter crescentus</i>

Copper (Cu) is potentially threatening for living organisms owing to its toxicity at high concentrations, requiring the onset of diverse detoxification strategies to maintain fitness. We previously showed that the environmental conditions modulate the response of the oligotrophic alphaproteobacterium <i>Caulobacter crescentus</i> to Cu excess. In the present study, we investigated the role of the Fe-importing TonB-dependent receptor (TBDR) CciT and its partner, CciO, a 2-oxoglutarate/Fe²⁺-dependent oxygenase, in Cu resistance. CciT is specifically involved in Cu resistance in both rich and poor media. Using inductively coupled plasma optical emission spectrometry, we found that under Cu stress, the cellular Cu content is reduced by overexpression of <i>cciT</i>, while the Fe content increases. Mutations of the three known Fe-importing TBDRs reveal that CciT is the primary Fe importer in these conditions and the only TBDR required for Cu resistance. In addition, the extracellular Fe concentration is positively correlated with the cellular Fe content and negatively correlated with the cellular Cu content, resulting in the protection of the cells against Cu excess. The operon organization of <i>cciT</i> and <i>cciO</i> is highly conserved across bacteria, indicating a functional link between the two proteins. Deletion of <i>cciT</i>, <i>cciO</i>, or both genes leads to similar Cu sensitivity. Catalytic mutations in CciT and CciO also result in Cu sensitivity. While CciO is not required for Cu and Fe transport, its precise function remains unknown. Overall, this study provides new insights into the role of Fe uptake in Cu resistance, emphasizing the critical influence of environmental conditions on bacterial physiology.IMPORTANCECopper is an essential metal for many living organisms, as it helps to drive crucial chemical reactions. However, when present in excess, copper turns toxic due to its high reactivity with biological molecules. Bacteria may encounter excess copper in various environments, such as polluted soils, agricultural copper treatments, and within the vacuoles of infected macrophages. In this study, we investigated the copper response in the environmental bacterium <i>Caulobacter crescentus</i>. Our findings reveal that environmental iron levels play a critical role in copper resistance, as increased iron prevents cellular copper accumulation and toxicity. We identified two essential proteins, CciT and CciO, that are involved in iron transport, providing protection against copper excess.