title: Overexpression of fetA (ybbL) and fetB (ybbM), Encoding an Iron Exporter, Enhances Resistance to Oxidative Stress in Escherichia coli.
abstract: Reactive oxygen species are generated by redox reactions and the Fenton reaction of H2O2 and iron that generates the hydroxyl radical that causes severe DNA, protein, and lipid damage. We screened Escherichia coli genomic libraries to identify a fragment, containing cueR, ybbJ, qmcA, ybbL, and ybbM, which enhanced resistance to H2O2 stress. We report that the ΔybbL and ΔybbM strains are more susceptible to H2O2 stress than the parent strain and that ybbL and ybbM overexpression overcomes H2O2 sensitivity. The ybbL and ybbM genes are predicted to code for an ATP-binding cassette metal transporter, and we demonstrate that YbbM is a membrane protein. We investigated various metals to identify iron as the likely substrate of this transporter. We propose the gene names fetA and fetB (for Fe transport) and the gene product names FetA and FetB. FetAB allows for increased resistance to oxidative stress in the presence of iron, revealing a role in iron homeostasis. We show that iron overload coupled with H2O2 stress is abrogated by fetA and fetB overexpression in the parent strain and in the Δfur strain, where iron uptake is deregulated. Furthermore, we utilized whole-cell electron paramagnetic resonance to show that intracellular iron levels in the Δfur strain are decreased by 37% by fetA and fetB overexpression. Combined, these findings show that fetA and fetB encode an iron exporter that has a role in enhancing resistance to H2O2-mediated oxidative stress and can minimize oxidative stress under conditions of iron overload and suggest that FetAB facilitates iron homeostasis to decrease oxidative stress.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
authors.forename: Sergios A
authors.initials: SA
authors.lastname: Nicolaou
authors.forename: Alan G
authors.initials: AG
authors.lastname: Fast
authors.forename: Eiko
authors.initials: E
authors.lastname: Nakamaru-Ogiso
authors.forename: Eleftherios T
authors.initials: ET
authors.lastname: Papoutsakis
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D006861:Hydrogen Peroxide / Q000633:toxicity; D007501:Iron / Q000378:metabolism*; D018384:Oxidative Stress
chemical_list: D001425:Bacterial Outer Membrane Proteins; C093120:FrpB protein, bacteria; D006861:Hydrogen Peroxide; D007501:Iron
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