Transcriptional regulation of ferric citrate transport in Escherichia coli K-12. Fecl belongs to a new subfamily of sigma 70-type factors that respond to extracytoplasmic stimuli.
Transcription of the ferric citrate transport system of Escherichia coli K-12 is repressed by Fe(2+)-Fur and activated by ferric citrate. Ferric citrate does not have to enter the cytoplasm; it initiates a signal transduction mechanism by binding to the outer membrane receptor FecA. Presumably, a conformational change is transmitted in a TonB-dependent manner to the FecR protein. FecR activates FecI, and FecI activates transcription of the fecABCDE transport genes. In this communication, FecI was isolated after cloning fecI downstream of an ideal ribosome-binding site. Overexpressed FecI formed inclusion bodies which were solubilized and purified in active form using a mild detergent. FecI, in conjunction with RNA polymerase core enzyme, directed transcription from the fecA promoter in an in vitro run-off transcription assay. Furthermore, FecI retarded the electrophoretic mobility of a specific 75 bp DNA fragment located upstream of fecA. An in vivo competition experiment between the fecA promoters of wild-type and mutant strains identified the nucleotide positions 2747, 2749, 2751 and 2753, located within the 75 bp fragment, as important for FecI-induced transcription. Mobility band shift of fecA promoter DNA caused by cell lysates required growth of cells in the presence of ferric citrate and expression of FecA, FecI and FecR. These data support the previous assignment of FecI, based on sequence homologies, to a new subfamily of eubacterial RNA polymerase sigma 70 factors that respond to extra-cytoplasmic stimuli and regulate extracytoplasmic functions.
Angerer
A
A
Universität Tübingen, Germany.
Enz
S
S
Ochs
M
M
Braun
V
V
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005290:Ferric Compounds / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D016623:Ion Pumps; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011956:Receptors, Cell Surface; D012808:Sigma Factor / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C028589:FecA protein, E coli; C490539:FecI protein, E coli; D005290:Ferric Compounds; D016623:Ion Pumps; D011956:Receptors, Cell Surface; D012808:Sigma Factor; C069970:FecB protein, E coli; C025314:ferric citrate; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases