Abstract
Escherichia coli fhuF mutants, a sufS::MudI mutant, and a sufD::MudI mutant were found to have the same phenotype: the inability to use ferrioxamine B as an iron source in a plate assay. In addition, the sufS and sufD genes were shown to be regulated by the iron-dependent Fur repressor. Sequence analysis revealed that the sufS open reading frame corresponds to orf f406. The protein SufS belongs to the family of NifS-like proteins, which supply sulfur for [Fe-S] centers. The protein FhuF contains a [2Fe-2S] center. A mutation in the upstream sufD gene (orf f423) caused the same phenotype. The T7 expression system and a His tag allow the isolation in good yield of the FhuF protein from a wild-type strain. In contrast, overproduction of the protein in a DeltasufD strain failed. Radioactive labeling of N-His-FhuF with [35S]methionine showed that the protein was unstable in the DeltasufD mutant.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Outer Membrane Proteins
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Bacterial Proteins / genetics*
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism*
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Bacterial Proteins / physiology
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Deferoxamine / metabolism
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Escherichia coli / drug effects
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Escherichia coli / genetics*
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Ferric Compounds / metabolism
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Gene Expression Regulation, Bacterial / drug effects
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Genes, Reporter / genetics
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Iron / pharmacology
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Iron-Binding Proteins
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Iron-Sulfur Proteins / genetics
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Iron-Sulfur Proteins / isolation & purification
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Iron-Sulfur Proteins / metabolism*
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Molecular Sequence Data
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Mutagenesis, Insertional
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Mutation
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Open Reading Frames / genetics
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Operon / genetics
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Periplasmic Binding Proteins
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Physical Chromosome Mapping
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Promoter Regions, Genetic / genetics
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Repressor Proteins / metabolism
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Temperature
Substances
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Bacterial Outer Membrane Proteins
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Bacterial Proteins
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Ferric Compounds
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Iron-Binding Proteins
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Iron-Sulfur Proteins
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Periplasmic Binding Proteins
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Recombinant Fusion Proteins
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Repressor Proteins
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ferric uptake regulating proteins, bacterial
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nifS protein, Bacteria
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ferrioxamine B
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Iron
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Deferoxamine