title: Exploring RNA polymerase regulation by NMR spectroscopy.
abstract: RNA synthesis is a central process in all organisms, with RNA polymerase (RNAP) as the key enzyme. Multisubunit RNAPs are evolutionary related and are tightly regulated by a multitude of transcription factors. Although Escherichia coli RNAP has been studied extensively, only little information is available about its dynamics and transient interactions. This information, however, are crucial for the complete understanding of transcription regulation in atomic detail. To study RNAP by NMR spectroscopy we developed a highly efficient procedure for the assembly of active RNAP from separately expressed subunits that allows specific labeling of the individual constituents. We recorded [(1)H,(13)C] correlation spectra of isoleucine, leucine, and valine methyl groups of complete RNAP and the separately labeled β' subunit within reconstituted RNAP. We further produced all RNAP subunits individually, established experiments to determine which RNAP subunit a certain regulator binds to, and identified the β subunit to bind NusE.
authors.affiliation: Lehrstuhl Biopolymere und Forschungszentrum für Bio-Makromoleküle, Universität Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Drögemüller
authors.affiliation: 1] [2] Lehrstuhl Biopolymere und Forschungszentrum für Bio-Makromoleküle, Universität Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Strauß
authors.affiliation: Lehrstuhl Biopolymere und Forschungszentrum für Bio-Makromoleküle, Universität Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
authors.forename: Kristian
authors.initials: K
authors.lastname: Schweimer
authors.affiliation: Lehrstuhl Biopolymere und Forschungszentrum für Bio-Makromoleküle, Universität Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
authors.forename: Birgitta M
authors.initials: BM
authors.lastname: Wöhrl
authors.affiliation: Lehrstuhl Biopolymere und Forschungszentrum für Bio-Makromoleküle, Universität Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
authors.forename: Stefan H
authors.initials: SH
authors.lastname: Knauer
authors.affiliation: Lehrstuhl Biopolymere und Forschungszentrum für Bio-Makromoleküle, Universität Bayreuth, Universitätsstraße 30, 95447 Bayreuth, Germany.
authors.forename: Paul
authors.initials: P
authors.lastname: Rösch
mesh_terms: D001665:Binding Sites; D066241:Carbon-13 Magnetic Resonance Spectroscopy; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011485:Protein Binding; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D066244:Proton Magnetic Resonance Spectroscopy; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D012269:Ribosomal Proteins; D014157:Transcription Factors; C029537:ribosomal protein S10; D012321:DNA-Directed RNA Polymerases