Characterization of Escherichia coli SecA protein binding to a site on its mRNA involved in autoregulation.
In order to understand further the autogenous regulation of Escherichia coli secA translation, we have set up a purified system to study the binding of SecA protein to portions of its mRNA. Specific SecA protein-RNA binding was demonstrated by UV cross-linking, filter binding, and gel shift assays. Use of the filter binding assay allowed optimization of binding, which was influenced by Mg2+ and ATP concentrations, and a measurement of the affinity of this interaction. A nested series of RNAs lacking either 5' or 3' portions of geneX-secA sequences were used to localize the SecA protein binding site to sequences around the geneX-secA intergenic region. These studies imply that SecA protein directly regulates its own translation by a specific RNA binding activity that presumably blocks translational initiation.
Dolan
K M
KM
Department of Microbiology, State University of New York, Stony Brook 11794.
Oliver
D B
DB
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D006207:Half-Life; D008274:Magnesium / Q000494:pharmacology; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D012329:RNA, Bacterial / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000069816:SEC Translocation Channels; D000081416:SecA Proteins; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D000069816:SEC Translocation Channels; D000255:Adenosine Triphosphate; D004247:DNA; D000251:Adenosine Triphosphatases; D000081416:SecA Proteins; D008274:Magnesium