Interaction of FurA from Anabaena sp. PCC 7120 with DNA: a reducing environment and the presence of Mn(2+) are positive effectors in the binding to isiB and furA promoters.
The Fur (ferric uptake regulator) protein is a global regulator in most prokaryotes that controls a large number of genes. Fur is a classical repressor that uses ferrous iron as co-repressor and binds to specific DNA sequences (iron boxes) as a dimer. Three different genes coding for Fur homologues have been identified in Anabaena sp. PCC 7120. FurA controls the transcription of flavodoxin, the product of the isiB gene, and is moderately autoregulated. In this work, the promoter of the furA gene was defined and the FurA protected regions in the furA and isiB promoters were identified, showing that the binding sites for Anabaena FurA contain A/T-rich sequences with a variable arrangement compared to the conventional 19-base pair Fur consensus. The influence of different factors on the interaction between FurA and the promoters was evaluated in vitro. The affinity of FurA for the DNA targets was significantly affected by the redox status of this regulator and the presence of Mn(2+). The optimal binding conditions were observed in the presence of both Mn(2+) and DTT. Those results suggest that, in addition to iron availability, FurA-DNA interaction is modulated by redox conditions.
Hernández
J A
JA
Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Ciencias, Universidad de Zaragoza, Pedro Cerbuna 12, 50009 Zaragoza, Spain.
López-Gomollón
S
S
Muro-Pastor
A
A
Valladares
A
A
Bes
M T
MT
Peleato
M L
ML
Fillat
M F
MF
D017033:Anabaena / Q000737:chemistry*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004229:Dithiothreitol / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D005418:Flavodoxin / Q000235:genetics*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic / Q000502:physiology*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
D001426:Bacterial Proteins; D005418:Flavodoxin; D012097:Repressor Proteins; C068477:ferric uptake regulating proteins, bacterial; D008345:Manganese; D006861:Hydrogen Peroxide; D004229:Dithiothreitol