title: RNA-binding to archaeal RNA polymerase subunits F/E: a DEER and FRET study.
abstract: RNA polymerases (RNAP) carry out transcription, the first step in the highly regulated process of gene expression. RNAPs are complex multisubunit enzymes, which undergo extensive structural rearrangements during the transcription cycle (initiation-elongation-termination). They accommodate interactions with the nucleic acid scaffold of transcription complexes (template DNA, DNA/RNA hybrid, and nascent RNA) and interact with a plethora of transcription factors. Here we focused on the RNAP-F/E subcomplex, which forms a stable heterodimer that binds the nascent RNA and thereby stimulates the processivity of elongation complexes. We used the pulsed-EPR method DEER and fluorescence spectroscopy to probe for conformational changes within the F/E dimer. Our results demonstrate that, upon binding of RNA, F/E remains in a stable conformation, which suggests that it serves as a structurally rigid guiding rail for the growing RNA chain during transcription.
authors.affiliation: Institute of Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, United Kingdom.
authors.forename: Dina
authors.initials: D
authors.lastname: Grohmann
authors.forename: Daniel
authors.initials: D
authors.lastname: Klose
authors.forename: Johann P
authors.initials: JP
authors.lastname: Klare
authors.forename: Christopher W M
authors.initials: CW
authors.lastname: Kay
authors.forename: Heinz-Jürgen
authors.initials: HJ
authors.lastname: Steinhoff
authors.forename: Finn
authors.initials: F
authors.lastname: Werner
mesh_terms: D018360:Crystallography, X-Ray; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D004583:Electrons; D031541:Fluorescence Resonance Energy Transfer; D017017:Methanococcus / Q000201:enzymology*; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D014158:Transcription, Genetic
chemical_list: D021122:Protein Subunits; D012313:RNA; D012321:DNA-Directed RNA Polymerases