title: Localization of a sigma70 binding site on the N terminus of the Escherichia coli RNA polymerase beta' subunit.
abstract: The Escherichia coli genome encodes genes for seven different sigma subunit species while only having single genes for the alpha, beta, and beta' subunits that make up the RNA polymerase core enzyme. The various sigma factors compete for binding to the core enzyme, upon which they confer promoter DNA-specific transcription initiation to the polymerase. We have mapped a major interaction site between one of the sigma species, sigma70, and beta'. Using far-Western blotting analysis of chemically cleaved and genetically engineered protein fragments, we have identified a N-terminal fragment of beta' (residues 60-309) that could bind sigma70. We were able to more precisely map the interaction domain to amino acid residues 260-309 of beta' using nickel nitrilotriacetic acid co-immobilization assays.
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin, Madison, Wisconsin 53706, USA.
authors.forename: T M
authors.initials: TM
authors.lastname: Arthur
authors.forename: R R
authors.initials: RR
authors.lastname: Burgess
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D015151:Immunoblotting; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives; D009942:Organometallic Compounds; D010449:Peptide Mapping; D011485:Protein Binding; D012808:Sigma Factor / Q000378:metabolism*
chemical_list: D009942:Organometallic Compounds; D012808:Sigma Factor; C088321:nickel nitrilotriacetic acid; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases; D009571:Nitrilotriacetic Acid