|Entry||Database: EMDB / ID: 9301|
|Title||Human TFIID BC core|
|Map data||primary map|
|Sample||General transcription factor IID:|
|Source||Homo sapiens (human)|
|Method||single particle reconstruction / cryo EM / 10.3 Å resolution|
|Authors||Patel AB / Louder RK|
|Citation||Journal: Science / Year: 2018|
Title: Structure of human TFIID and mechanism of TBP loading onto promoter DNA.
Authors: Avinash B Patel / Robert K Louder / Basil J Greber / Sebastian Grünberg / Jie Luo / Jie Fang / Yutong Liu / Jeff Ranish / Steve Hahn / Eva Nogales
Abstract: The general transcription factor IID (TFIID) is a critical component of the eukaryotic transcription preinitiation complex (PIC) and is responsible for recognizing the core promoter DNA and ...The general transcription factor IID (TFIID) is a critical component of the eukaryotic transcription preinitiation complex (PIC) and is responsible for recognizing the core promoter DNA and initiating PIC assembly. We used cryo-electron microscopy (cryo-EM), chemical crosslinking-mass spectrometry (CX-MS) and biochemical reconstitution to determine the complete molecular architecture of TFIID and define the conformational landscape of TFIID in the process of TATA-box binding protein (TBP) loading onto promoter DNA. Our structural analysis revealed five structural states of TFIID in the presence of TFIIA and promoter DNA, showing that the initial binding of TFIID to the downstream promoter positions the upstream DNA and facilitates scanning of TBP for a TATA-box and the subsequent engagement of the promoter. Our findings provide a mechanistic model for the specific loading of TBP by TFIID onto the promoter.
|Date||Deposition: Nov 5, 2018 / Header (metadata) release: Nov 28, 2018 / Map release: Nov 28, 2018 / Last update: Dec 5, 2018|
|Structure viewer||EM map: |
Downloads & links
|File||emd_9301.map.gz (map file in CCP4 format, 95552 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.32 Å|
CCP4 map header:
-Entire General transcription factor IID
|Entire||Name: General transcription factor IID / Number of components: 1|
-Component #1: protein, General transcription factor IID
|Protein||Name: General transcription factor IID / Recombinant expression: No|
|Source||Species: Homo sapiens (human) / Strain: HeLa|
|Specimen||Specimen state: particle / Method: cryo EM|
|Sample solution||Specimen conc.: .2 mg/ml / pH: 7.9|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 277.15 K / Humidity: 100 % / Details: BT 4s; BF 15N.|
-Electron microscopy imaging
|Imaging||Microscope: FEI TITAN|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 4 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Cs: 2.7 mm / Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: GATAN LIQUID NITROGEN|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 103068|
|3D reconstruction||Resolution: 10.3 Å / Resolution method: FSC 0.143 CUT-OFF|
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