Domain analysis of the Edwardsiella tarda ferric uptake regulator.
Recent studies have shown that the ferric uptake regulator (Fur) of Edwardsiella tarda (Fur(Et)) shares high sequence identity with the Escherichia coli Fur (Fur(Ec)) at the N-terminal DNA-binding region. In the present study, the functional importance of the C-terminal region of Fur(Et) was investigated. It was found that Fur(Et) bearing deletion of the C-terminal 12 residues still possesses most of the repressor activity, whereas Fur(Et) bearing deletions of the C-terminal 16 and more than 16 residues are severely affected in activity. Domain swapping analyses indicated that the chimeric Fur proteins (Et75Ec73 and Et75Vh74) consisting of the N-terminal 1-75 region of Fur(Et) fused to the C-terminal 76-148 region of Fur(Ec) and the C-terminal 76-149 region of the Vibrio harveyi Fur (Fur(Vh)), respectively, are fully active. C92 of Fur(Ec) and C137 of Fur(Vh), which are functionally essential in Fur(Ec) and Fur(Vh), respectively, are also essential in Et75Ec73 and Et75Vh74, respectively. Further study identified an artificial Fur protein, EtMF54, which is composed of the N-terminal 49 residues of Fur(Et) and five artificial residues. Compared to Fur(Et), EtMF54 possesses partial Fur activity that is iron-dependent. These results (i) indicate that there exist certain functional/structural compatibilities among Fur(Et), Fur(Ec), and Fur(Vh) at the C-terminal region; (ii) provide insights to the potential location of the regulatory ion-binding site of Fur(Et).
Sun
Kun
K
Institute of Oceanology, Chinese Academy of Sciences, Qingdao, PR China.
Cheng
Shuang
S
Wang
Fang
F
Sun
Li
L
D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D020609:Edwardsiella tarda / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D007501:Iron / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D012097:Repressor Proteins / Q000737:chemistry*
D001426:Bacterial Proteins; D012097:Repressor Proteins; C068477:ferric uptake regulating proteins, bacterial; D007501:Iron