Three fur homologues from Anabaena sp. PCC7120: exploring reciprocal protein-promoter recognition.
DNA sequence analysis of the Anabaena sp. PCC7120 genome confirmed the presence of three open reading frames (all1691, all2473 and alr0957) containing the histidine-rich region characteristic of the Fur family. The genes coding for the three Fur proteins were cloned and overexpressed in Escherichia coli. The overexpression products, called FurA, FurB and FurC are only distantly related. The ability of the three recombinant proteins to bind iron-boxes identified in the three fur promoter regions was tested by electrophoretical mobility shift assays. FurA binds the three fur promoters with increased affinity in presence of metal. FurB also binds the three fur promoters, and its affinity is increased with DTT. FurC does not bind to furA or furB promoter regions or to its own promoter. However, FurC affects the ability of FurB and FurA to bind their target promoters.
Hernández
José A
JA
Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Ciencias, Universidad de Zaragoza, 50009 Zaragoza, Spain.
López-Gomollón
Sara
S
Bes
M Teresa
MT
Fillat
María F
MF
Peleato
M Luisa
ML
D017033:Anabaena / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004229:Dithiothreitol / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007501:Iron / Q000378:metabolism; D008670:Metals / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D008670:Metals; D011994:Recombinant Proteins; D012097:Repressor Proteins; C068477:ferric uptake regulating proteins, bacterial; D007501:Iron; D004229:Dithiothreitol