Surface signaling in ferric citrate transport gene induction: interaction of the FecA, FecR, and FecI regulatory proteins.
In Escherichia coli, transcription of the ferric citrate transport genes fecABCDE is controlled by a novel signal transduction mechanism that starts at the cell surface. Binding of ferric citrate to the outer membrane protein FecA initiates a signal that is transmitted by FecR across the cytoplasmic membrane into the cytoplasm where FecI, the sigma factor, is activated. Interaction between the signaling proteins was demonstrated by utilizing two methods. In in vitro binding assays, FecR that was His tagged at the N terminus [(His)(10)-FecR] and bound to a Ni-nitrilotriacetic acid agarose column was able to retain FecA, and FecR that was His tagged at the C terminus [FecR-(His)(6)] retained FecI on the column. An N-terminally truncated, induction-negative but transport-active FecA protein did not bind to (His)(10)-FecR. The in vivo assay involved the determination of the FecA, FecR, and FecI interacting domains with the bacterial two-hybrid Lex-based system. FecA(1-79) interacts with FecR(101-317) and FecR(1-85) interacts with FecI(1-173). These data clearly support a model that proposes interaction of the periplasmic N terminus of FecA with the periplasmic C-terminal portion of FecR and interaction of the cytoplasmic N terminus of FecR with FecI, which results in FecI activation.
Enz
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Mikrobiologie/Membranphysiologie, Universität Tübingen, D-72076 Tübingen, Germany.
Mahren
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Stroeher
U H
UH
Braun
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V
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001693:Biological Transport, Active; D002846:Chromatography, Affinity; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D005290:Ferric Compounds / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006639:Histidine / Q000378:metabolism; D008970:Molecular Weight; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism*; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C490539:FecI protein, E coli; D005290:Ferric Compounds; C489318:RecR protein, E coli; D012808:Sigma Factor; C065831:RecR protein, Bacteria; D006639:Histidine; C025314:ferric citrate; D011938:Rec A Recombinases