title: Complete architecture of the archaeal RNA polymerase open complex from single-molecule FRET and NPS.
abstract: The molecular architecture of RNAP II-like transcription initiation complexes remains opaque due to its conformational flexibility and size. Here we report the three-dimensional architecture of the complete open complex (OC) composed of the promoter DNA, TATA box-binding protein (TBP), transcription factor B (TFB), transcription factor E (TFE) and the 12-subunit RNA polymerase (RNAP) from Methanocaldococcus jannaschii. By combining single-molecule Förster resonance energy transfer and the Bayesian parameter estimation-based Nano-Positioning System analysis, we model the entire archaeal OC, which elucidates the path of the non-template DNA (ntDNA) strand and interaction sites of the transcription factors with the RNAP. Compared with models of the eukaryotic OC, the TATA DNA region with TBP and TFB is positioned closer to the surface of the RNAP, likely providing the mechanism by which DNA melting can occur in a minimal factor configuration, without the dedicated translocase/helicase encoding factor TFIIH.
authors: Biophysics Institute, Ulm University, Albert-Einstein-Allee 11, Ulm 89069, Germany.
authors: Julia
authors: J
authors: Nagy
authors: Institut für Physikalische und Theoretische Chemie-NanoBioSciences, Technische Universität Braunschweig, Hans-Sommer-Straße 10, 38106 Braunschweig, Germany.
authors: Dina
authors: D
authors: Grohmann
authors: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors: Alan C M
authors: AC
authors: Cheung
authors: Institut für Physikalische und Theoretische Chemie-NanoBioSciences, Technische Universität Braunschweig, Hans-Sommer-Straße 10, 38106 Braunschweig, Germany.
authors: Sarah
authors: S
authors: Schulz
authors: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors: Katherine
authors: K
authors: Smollett
authors: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors: Finn
authors: F
authors: Werner
authors: Biophysics Institute, Ulm University, Albert-Einstein-Allee 11, Ulm 89069, Germany.
authors: Jens
authors: J
authors: Michaelis
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism*; D019641:DNA, Archaeal / Q000737:chemistry / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000378:metabolism*; D019642:RNA, Archaeal / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D019642:RNA, Archaeal; D012321:DNA-Directed RNA Polymerases
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