Iron is an essential micronutrient, and bacteria make use of several strategies for regulating its levels. Among these, they utilize proteins that store iron (ferritins), which liberate the metal when present in low concentration, and control iron availability by regulating the expression of these proteins. In several bacteria, this regulation is mediated by iron containing-Fur, which acts directly or indirectly via a small regulatory RNA (sRNA). This work aims to study two ferritins and their regulatory mechanisms, including the identificationin C. crescentus of a functional equivalent to the small RNA RhyB from E. coli. The ferritin genes and the region containing the sRNA will be deleted, and the mutants phenotypes will be evaluated as to iron metabolism and oxidative stress response. Gene expression assays in different iron homeostasis conditions will be performed by cloning the promoters in front of a reporte lacZ gene. Expression of ferritins will be evaluated in the mutants for sRNA, fur and oxyR, which will allow defining the regulatory network for these genes. Expression of other genes important for iron metabolism will be assayed in the sRNA mutant and the role of each ferritin in iron accumulation will be determined.
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