The FecI extracytoplasmic-function sigma factor of Escherichia coli interacts with the beta' subunit of RNA polymerase.
Transcription of the ferric citrate transport system of Escherichia coli K-12 is mediated by the extracytoplasmic-function (ECF) sigma factor FecI, which is activated by ferric citrate in the growth medium. By using a bacterial two-hybrid system, it was shown in vivo that FecI binds to the beta' subunit of RNA polymerase. The inactive mutant protein FecI(K155E) displayed reduced binding to beta', and small deletions along the entire FecI protein led to total impairment of beta' binding. In vitro, FecI was retained on Ni(2+)-nitrilotriacetic acid agarose loaded with a His-tagged beta'(1-313) fragment and coeluted with beta'(1-313). Binding of FecI to beta' and beta'(1-313) was enhanced by FecR(1-85), which represents the cytoplasmic portion of the FecR protein that transmits the inducing signal across the cytoplasmic membrane. Interaction of FecR with FecI was demonstrated by showing that isolated FecR inhibited degradation of FecI by trypsin. This is the first demonstration of binding of an ECF sigma factor of the FecI type to the beta' subunit of RNA polymerase and of binding being enhanced by the protein that activates the ECF sigma factor.
Mahren
Susanne
S
Mikrobiologie/Membranphysiologie, Universität Tübingen, D-72076 Tübingen, Germany.
Braun
Volkmar
V
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D026901:Membrane Transport Proteins; D009154:Mutation; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C490539:FecI protein, E coli; C093541:FecR protein, E coli; D026901:Membrane Transport Proteins; D012808:Sigma Factor; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit