Experimental and computational characterization of the ferric uptake regulator from Aliivibrio salmonicida (Vibrio salmonicida).
The Ferric uptake regulator (Fur) is a global transcription factor that affects expression of bacterial genes in an iron-dependent fashion. Although the Fur protein and its iron-responsive regulon are well studied, there are still important questions that remain to be answered. For example, the consensus Fur binding site also known as the "Fur box" is under debate, and it is still unclear which Fur residues directly interact with the DNA. Our long-term goal is to dissect the biological roles of Fur in the development of the disease cold-water vibriosis, which is caused by the psychrophilic bacteria Aliivibrio salmonicida (also known as Vibrio salmonicida). Here, we have used experimental and computational methods to characterise the Fur protein from A. salmonicida (AS-Fur). Electrophoretic mobility shift assays show that AS-Fur binds to the recently proposed vibrio Fur box consensus in addition to nine promoter regions that contain Fur boxes. Binding appears to be dependent on the number of Fur boxes, and the predicted "strength" of Fur boxes. Finally, structure modeling and molecular dynamics simulations provide new insights into potential AS-Fur-DNA interactions.
Pedersen
Hege Lynum
HL
Department of Molecular Biotechnology, Institute of Medical Biology, Faculty of Medicine, University of Tromsø, Tromsø, N-9037, Norway.
Ahmad
Rafi
R
Riise
Ellen Kristin
EK
Leiros
Hanna-Kirsti Schrøder
HK
Hauglid
Stefan
S
Espelid
Sigrun
S
Brandsdal
Bjøn Olav
BO
Leiros
Ingar
I
Willassen
Nils-Peder
NP
Haugen
Peik
P
D044165:Aliivibrio salmonicida / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D004269:DNA, Bacterial / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005290:Ferric Compounds / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005290:Ferric Compounds; D012097:Repressor Proteins; C068477:ferric uptake regulating proteins, bacterial