|Entry||Database: EMDB / ID: EMD-7998|
|Title||RNA Pol II(G) focus refinement|
|Biological species||Homo sapiens (human)|
|Method||single particle reconstruction / cryo EM / Resolution: 4.3 Å|
|Authors||Yu X / Jishage M|
|Citation||Journal: Nat. Struct. Mol. Biol. / Year: 2018|
Title: Architecture of Pol II(G) and molecular mechanism of transcription regulation by Gdown1.
Authors: Miki Jishage / Xiaodi Yu / Yi Shi / Sai J Ganesan / Wei-Yi Chen / Andrej Sali / Brian T Chait / Francisco J Asturias / Robert G Roeder /
Abstract: Tight binding of Gdown1 represses RNA polymerase II (Pol II) function in a manner that is reversed by Mediator, but the structural basis of these processes is unclear. Although Gdown1 is ...Tight binding of Gdown1 represses RNA polymerase II (Pol II) function in a manner that is reversed by Mediator, but the structural basis of these processes is unclear. Although Gdown1 is intrinsically disordered, its Pol II interacting domains were localized and shown to occlude transcription factor IIF (TFIIF) and transcription factor IIB (TFIIB) binding by perfect positioning on their Pol II interaction sites. Robust binding of Gdown1 to Pol II is established by cooperative interactions of a strong Pol II binding region and two weaker binding modulatory regions, thus providing a mechanism both for tight Pol II binding and transcription inhibition and for its reversal. In support of a physiological function for Gdown1 in transcription repression, Gdown1 co-localizes with Pol II in transcriptionally silent nuclei of early Drosophila embryos but re-localizes to the cytoplasm during zygotic genome activation. Our study reveals a self-inactivation through Gdown1 binding as a unique mode of repression in Pol II function.
|Date||Deposition: Jun 11, 2018 / Header (metadata) release: Jul 25, 2018 / Map release: Jun 12, 2019 / Update: Jun 12, 2019|
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_7998.map.gz / Format: CCP4 / Size: 42.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.31 Å|
|Symmetry||Space group: 1|
CCP4 map header:
|Entire||Name: hPolII_Gdown1 / Number of components: 1|
-Component #1: protein, hPolII_Gdown1
|Protein||Name: hPolII_Gdown1 / Recombinant expression: No|
|Source||Species: Homo sapiens (human)|
|Specimen||Specimen state: Cell / Method: cryo EM|
|Sample solution||pH: 7.5|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 40 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Cs: 2.7 mm / Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 35026|
|3D reconstruction||Resolution: 4.3 Å / Resolution method: FSC 0.143 CUT-OFF|
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