Autoregulation of Helicobacter pylori Fur revealed by functional analysis of the iron-binding site.
The ferric uptake regulator protein Fur regulates iron-dependent gene expression in bacteria. In Helicobacter pylori it has been shown to regulate iron-activated and iron-repressed genes. In this study, we show that H. pylori Fur protein regulates transcription from its own sigma 80 promoter P fur in response to iron. Footprinting analysis shows that Fur binds at three distinct operators at P fur overlapping and proximal to the promoter elements. Site-directed mutagenesis of the proposed iron-binding site of the protein results in derepression of P fur and the loss of iron regulation. In vivo oligomerization assays reveals that the C-terminus of Fur is necessary for multimerization of the protein and that the mutations do not affect this activity. Molecular and phenotypic analysis of the mutant proteins provides evidence that the iron-binding site controls the specific affinity of Fur for the operators at P fur and hence its repressive ability. In summary, the data presented are consistent with a model in which Fur acts as a rheostat of transcription to autoregulate its own expression in response to iron, which in turn controls expression of iron-induced and iron-repressed genes, providing maintenance of homeostasis.
Delany
Isabel
I
Department of Molecular Biology, Siena, italy.
Spohn
Gunther
G
Pacheco
Ana-Beatriz F
AB
Ieva
Raffaele
R
Alaimo
Cristina
C
Rappuoli
Rino
R
Scarlato
Vincenzo
V
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D019281:Dimerization; D005816:Genetic Complementation Test; D016480:Helicobacter pylori / Q000235:genetics / Q000378:metabolism*; D006706:Homeostasis; D007501:Iron / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010641:Phenotype; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D012097:Repressor Proteins; C068477:ferric uptake regulating proteins, bacterial; D007501:Iron