1887

Abstract

A homologue of the gene was identified in and characterized. The predicted amino acid sequence of the cloned gene was found to be highly similar to other members of the Fur family of transcriptional regulators. The gene was shown to partially complement an mutant. Purified Fur bound specifically to a 19 bp DNA sequence homologous to the Fur box in a metal-dependent manner. Analysis of the available genome data identified a number of genes which contain predicted Fur box sequences in the promoter region. Many of these genes are predicted to play a role in bacterial iron uptake and metabolism, but several have also been implicated as having a role in virulence. Fur and iron regulation of a siderophore biosynthesis operon was confirmed in a -galactosidase assay. A null strain was constructed by allelic replacement of the chromosomal gene with a copy disrupted with a kanamycin resistance cassette. The Δ mutant was found to constitutively express siderophores, to accumulate iron intracellularly to a level approximately threefold greater than the wild-type, and to be hypersensitive to hydrogen peroxide. In an insect infection model, the virulence of the null strain was found to be significantly attenuated, highlighting the essential role played by Fur in the virulence of this pathogen.

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2005-02-01
2024-12-02
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