Efficient transcriptional antitermination from the Escherichia coli cytoplasmic membrane.
The BglG protein is a transcriptional antiterminator acting within the beta-glucoside operon of Escherichia coli by binding to a specific sequence motif in the growing mRNA. Binding of BglG prevents formation of the terminator stem-loop structure, thereby causing the RNA polymerase to continue transcription. Activity of BglG is modulated in a complex way by antagonistically acting phosphorylations in response to the availability of beta-glucosidic substrates and to the catabolic state of the cell. The enzymes responsible for these phosphorylations are members of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) that represents a central carbohydrate uptake and signal transduction system. As these enzymes are believed to all form higher-order complexes associated with the cytoplasmic membrane, we tested whether or not BglG would remain active when artificially anchored to its presumptive site of regulation, the inner membrane. We show that the membrane-anchored protein indeed efficiently catalyzes transcriptional antitermination. Moreover, the membrane-attached BglG remains regulated by the PTS. Thus, a membrane-bound regulatory RNA binding protein can potentially interact fast enough with its target within the nascent transcript and cause the transcriptional machinery to proceed, before transcriptional termination would occur. Consequently, there is no principal necessity for an RNA-binding transcriptional regulator like BglG to leave the inner membrane, a potential regulatory site, and migrate to the site of transcription, the nucleoid.
Görke
B
B
Institut für Biologie III, Universität, Schänzlestr. 1, D-79104 Freiburg, Germany.
Rak
B
B
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D017104:Bacteriophage M13 / Q000235:genetics; D002213:Capsid / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology* / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism; D010766:Phosphorylation; D021381:Protein Transport; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic / Q000235:genetics*
D001426:Bacterial Proteins; D036022:Capsid Proteins; D008565:Membrane Proteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C064872:antiterminator proteins, Bacteria; C025668:coat protein, Bacteriophage M13; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System