A single amino acid change in AngR, a protein encoded by pJM1-like virulence plasmids, results in hyperproduction of anguibactin.
The siderophore anguibactin is produced in vivo in a diffusible form and is an important factor in the virulence of Vibrio anguillarum. The natural isolate V. anguillarum 531A is a hyperproducer of anguibactin when compared with the prototype strain V. anguillarum 775. The angR gene was found to be responsible for this difference in levels of anguibactin produced. Nucleotide sequence analysis showed that the angR531A differed in a single nucleotide from the angR775 present in the prototype plasmid pJM1. This nucleotide substitution resulted in a change in amino acid 267 from His in strain 775 to Asn in strain 531A. This amino acid is located in a region between one of the two helix-turn-helix domains and the neighboring leucine zipper. Mutations to replace His with either Leu or Gln, generated by site-directed mutagenesis, in amino acid 267 resulted in strains for which the MIC of the iron chelator ethylenediamine di(o-hydroxyphenyl) acetic acid were lower than for the proptotype 775 but higher than for iron uptake-deficient strains. In addition to its transcriptional activating function, AngR also complemented a mutation in the Escherichia coli entE gene, which encodes the enterobactin biosynthetic enzyme 2,3-dihydroxybenzoate-AMP ligase. Therefore, AngR may also function in V. anguillarum as an EntE-like enzyme for the biosynthesis of anguibactin.
Tolmasky
M E
ME
Department of Microbiology and Immunology, School of Medicine, Oregon Health Sciences University, Portland 97201-3098.
Actis
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LA
Crosa
J H
JH
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D004268:DNA-Binding Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D010455:Peptides; D010957:Plasmids; D017262:Siderophores / Q000096:biosynthesis*; D014157:Transcription Factors; D014733:Vibrio / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D014774:Virulence
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D010455:Peptides; D017262:Siderophores; D014157:Transcription Factors; C063474:angR protein, Vibrio anguillarum; C049469:anguibactin