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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-8133 | |||||||||||||||
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Title | Human holo-PIC in the initial transcribing state | |||||||||||||||
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![]() | initiation / RNA polymerase II / human / TRANSCRIPTION / TRANSFERASE-DNA complex | |||||||||||||||
Function / homology | ![]() LRR domain binding / microfibril binding / MMXD complex / core TFIIH complex portion of holo TFIIH complex / Cytosolic iron-sulfur cluster assembly / : / central nervous system myelin formation / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / positive regulation of core promoter binding ...LRR domain binding / microfibril binding / MMXD complex / core TFIIH complex portion of holo TFIIH complex / Cytosolic iron-sulfur cluster assembly / : / central nervous system myelin formation / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / positive regulation of core promoter binding / RNA polymerase II core complex assembly / meiotic sister chromatid cohesion / positive regulation of mitotic recombination / hair cell differentiation / hair follicle maturation / nucleotide-excision repair factor 3 complex / transcription factor TFIIE complex / RNA polymerase transcription factor SL1 complex / regulation of transcription by RNA polymerase I / nucleotide-excision repair, preincision complex assembly / phosphatase activator activity / CAK-ERCC2 complex / embryonic cleavage / RPAP3/R2TP/prefoldin-like complex / RNA polymerase III general transcription initiation factor activity / TFIIF-class transcription factor complex binding / UV protection / transcriptional start site selection at RNA polymerase II promoter / RNA polymerase I core promoter sequence-specific DNA binding / transcription factor TFIIF complex / DNA 5'-3' helicase / Cytosolic sensors of pathogen-associated DNA / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Polymerase III Transcription Initiation From Type 3 Promoter / transcription factor TFIIA complex / RNA Polymerase III Abortive And Retractive Initiation / G protein-coupled receptor internalization / female germ cell nucleus / transcription factor TFIIH core complex / transcription factor TFIIH holo complex / male pronucleus / female pronucleus / germinal vesicle / nuclear thyroid hormone receptor binding / RNA polymerase II general transcription initiation factor binding / Abortive elongation of HIV-1 transcript in the absence of Tat / FGFR2 alternative splicing / RNA Polymerase I Transcription Termination / regulation of mitotic cell cycle phase transition / transcription preinitiation complex / MicroRNA (miRNA) biogenesis / 3'-5' DNA helicase activity / DNA 3'-5' helicase / Viral Messenger RNA Synthesis / Signaling by FGFR2 IIIa TM / protein acetylation / transcription factor TFIID complex / RNA polymerase II general transcription initiation factor activity / cell division site / spinal cord development / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / hematopoietic stem cell proliferation / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / erythrocyte maturation / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / mRNA Capping / HIV Transcription Initiation / RNA Polymerase II HIV Promoter Escape / Transcription of the HIV genome / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Initiation And Promoter Clearance / bone mineralization / acetyltransferase activity / mRNA Splicing - Minor Pathway / RNA polymerase II complex binding / PIWI-interacting RNA (piRNA) biogenesis / viral transcription / ATPase activator activity / aryl hydrocarbon receptor binding / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / transcription by RNA polymerase I / intrinsic apoptotic signaling pathway by p53 class mediator / DNA topological change / TFIIB-class transcription factor binding / RNA Polymerase I Transcription Initiation / nuclear-transcribed mRNA catabolic process / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / Processing of Capped Intron-Containing Pre-mRNA / positive regulation of translational initiation / transcription by RNA polymerase III / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / RNA polymerase II transcribes snRNA genes / hematopoietic stem cell differentiation Similarity search - Function | |||||||||||||||
Biological species | ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 7.9 Å | |||||||||||||||
![]() | He Y / Yan C | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Near-atomic resolution visualization of human transcription promoter opening. Authors: Yuan He / Chunli Yan / Jie Fang / Carla Inouye / Robert Tjian / Ivaylo Ivanov / Eva Nogales / ![]() Abstract: In eukaryotic transcription initiation, a large multi-subunit pre-initiation complex (PIC) that assembles at the core promoter is required for the opening of the duplex DNA and identification of the ...In eukaryotic transcription initiation, a large multi-subunit pre-initiation complex (PIC) that assembles at the core promoter is required for the opening of the duplex DNA and identification of the start site for transcription by RNA polymerase II. Here we use cryo-electron microscropy (cryo-EM) to determine near-atomic resolution structures of the human PIC in a closed state (engaged with duplex DNA), an open state (engaged with a transcription bubble), and an initially transcribing complex (containing six base pairs of DNA-RNA hybrid). Our studies provide structures for previously uncharacterized components of the PIC, such as TFIIE and TFIIH, and segments of TFIIA, TFIIB and TFIIF. Comparison of the different structures reveals the sequential conformational changes that accompany the transition from each state to the next throughout the transcription initiation process. This analysis illustrates the key role of TFIIB in transcription bubble stabilization and provides strong structural support for a translocase activity of XPB. | |||||||||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 1.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 55.2 KB 55.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 6.9 KB | Display | ![]() |
Images | ![]() | 155.6 KB | ||
Filedesc metadata | ![]() | 14.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 374.1 KB | Display | ![]() |
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Full document | ![]() | 373.6 KB | Display | |
Data in XML | ![]() | 9.1 KB | Display | |
Data in CIF | ![]() | 11.7 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5iy8MC ![]() 8131C ![]() 8132C ![]() 8134C ![]() 8135C ![]() 8136C ![]() 8137C ![]() 8138C ![]() 5ivwC ![]() 5iy6C ![]() 5iy7C ![]() 5iy9C ![]() 5iyaC ![]() 5iybC ![]() 5iycC ![]() 5iydC C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | None | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.62 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : Human holo-PIC in the initial transcribing state
+Supramolecule #1: Human holo-PIC in the initial transcribing state
+Supramolecule #2: RNA polymerase II
+Supramolecule #3: General transcription factors
+Supramolecule #4: Super core promoter
+Macromolecule #1: DNA-directed RNA polymerase II subunit RPB1
+Macromolecule #2: DNA-directed RNA polymerase II subunit RPB2
+Macromolecule #3: DNA-directed RNA polymerase II subunit RPB3
+Macromolecule #4: DNA-directed RNA polymerase II subunit RPB4
+Macromolecule #5: DNA-directed RNA polymerase II subunit RPB5
+Macromolecule #6: DNA-directed RNA polymerase II subunit RPB6
+Macromolecule #7: DNA-directed RNA polymerase II subunit RPB7
+Macromolecule #8: DNA-directed RNA polymerase II subunit RPB8
+Macromolecule #9: DNA-directed RNA polymerase II subunit RPB9
+Macromolecule #10: DNA-directed RNA polymerase II subunit RPB10
+Macromolecule #11: DNA-directed RNA polymerase II subunit RPB11-a
+Macromolecule #12: DNA-directed RNA polymerase II subunit RPB12
+Macromolecule #13: Transcription initiation factor IIB
+Macromolecule #14: Transcription initiation factor IIA subunit 1
+Macromolecule #15: Transcription initiation factor IIA subunit 2
+Macromolecule #16: TATA-box-binding protein
+Macromolecule #17: General transcription factor IIE subunit 1
+Macromolecule #18: Transcription initiation factor IIE subunit beta
+Macromolecule #19: General transcription factor IIF subunit 1
+Macromolecule #20: General transcription factor IIF subunit 2
+Macromolecule #21: Transcription elongation factor A protein 1
+Macromolecule #22: TFIIH basal transcription factor complex helicase XPB subunit
+Macromolecule #23: TFIIH basal transcription factor complex helicase XPD subunit
+Macromolecule #24: General transcription factor IIH subunit 2
+Macromolecule #25: General transcription factor IIH subunit 5
+Macromolecule #26: General transcription factor IIH subunit 4
+Macromolecule #27: General transcription factor IIH subunit 3
+Macromolecule #28: SCP-X
+Macromolecule #29: SCP-Y
+Macromolecule #30: MAGNESIUM ION
+Macromolecule #31: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.3 mg/mL |
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Buffer | pH: 7.9 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV Details: Blot for 4 seconds before plunging into liquid ethane (FEI VITROBOT MARK IV).. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 42.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 4.0 µm / Nominal defocus min: 2.0 µm / Nominal magnification: 27500 |
Sample stage | Specimen holder model: GATAN 626 SINGLE TILT LIQUID NITROGEN CRYO TRANSFER HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Protocol: FLEXIBLE FIT |
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Output model | ![]() PDB-5iy8: |