RNA polymerase I-specific transcription initiation factor RRN6 / DNA-directed RNA polymerase I subunit RPA49 / DNA-directed RNA polymerase I subunit RPA14 / un:p47006: / DNA-directed RNA polymerase I subunit RPA43 / RNA polymerase I-specific transcription initiation factor RRN7 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / DNA-directed RNA polymerases I, II, and III subunit RPABC1 / DNA-directed RNA polymerase I subunit RPA12 / DNA-directed RNA polymerases I and III subunit RPAC2 ...RNA polymerase I-specific transcription initiation factor RRN6 / DNA-directed RNA polymerase I subunit RPA49 / DNA-directed RNA polymerase I subunit RPA14 / un:p47006: / DNA-directed RNA polymerase I subunit RPA43 / RNA polymerase I-specific transcription initiation factor RRN7 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / DNA-directed RNA polymerases I, II, and III subunit RPABC1 / DNA-directed RNA polymerase I subunit RPA12 / DNA-directed RNA polymerases I and III subunit RPAC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC5 / DNA-directed RNA polymerase I subunit RPA135 / DNA-directed RNA polymerases I, II, and III subunit RPABC3 / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerase I subunit RPA190 / DNA-directed RNA polymerases I and III subunit RPAC1 / RNA polymerase I-specific transcription initiation factor RRN11
Biological species
Saccharomyces cerevisiae S288c (yeast)
Method
single particle reconstruction / cryo EM / Resolution: 6.9 Å
Journal: Elife / Year: 2017 Title: Structural mechanism of ATP-independent transcription initiation by RNA polymerase I. Authors: Yan Han / Chunli Yan / Thi Hoang Duong Nguyen / Ashleigh J Jackobel / Ivaylo Ivanov / Bruce A Knutson / Yuan He / Abstract: Transcription initiation by RNA Polymerase I (Pol I) depends on the Core Factor (CF) complex to recognize the upstream promoter and assemble into a Pre-Initiation Complex (PIC). Here, we solve a ...Transcription initiation by RNA Polymerase I (Pol I) depends on the Core Factor (CF) complex to recognize the upstream promoter and assemble into a Pre-Initiation Complex (PIC). Here, we solve a structure of Pol I-CF-DNA to 3.8 Å resolution using single-particle cryo-electron microscopy. The structure reveals a bipartite architecture of Core Factor and its recognition of the promoter from -27 to -16. Core Factor's intrinsic mobility correlates well with different conformational states of the Pol I cleft, in addition to the stabilization of either Rrn7 N-terminal domain near Pol I wall or the tandem winged helix domain of A49 at a partially overlapping location. Comparison of the three states in this study with the Pol II system suggests that a ratchet motion of the Core Factor-DNA sub-complex at upstream facilitates promoter melting in an ATP-independent manner, distinct from a DNA translocase actively threading the downstream DNA in the Pol II PIC.
History
Deposition
Jun 16, 2017
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Header (metadata) release
Jul 5, 2017
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Map release
Jul 5, 2017
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Update
Jul 18, 2018
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Current status
Jul 18, 2018
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Electron dose: 17.2 e/Å2 / Illumination mode: FLOOD BEAM
Lens
Magnification: 15000. X (nominal) / Cs: 2 mm / Imaging mode: BRIGHT FIELD
Specimen Holder
Model: GATAN LIQUID NITROGEN
Camera
Detector: GATAN K2 SUMMIT (4k x 4k)
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Image processing
Processing
Method: single particle reconstruction / Applied symmetry: C1 (asymmetric) / Number of projections: 43843
3D reconstruction
CTF correction: CTF amplitude correction was performed following 3D auto refinement in relion. Resolution: 6.9 Å / Resolution method: FSC 0.143 CUT-OFF
FSC plot (resolution estimation)
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Atomic model buiding
Modeling #1
Refinement space: REAL
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