+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-8134 | |||||||||||||||
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Title | Human holo-PIC in the initial transcribing state (no IIS) | |||||||||||||||
Map data | None | |||||||||||||||
Sample |
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Keywords | initiation / RNA polymerase II / human / TRANSCRIPTION / TRANSFERASE-DNA-RNA complex | |||||||||||||||
Function / homology | Function and homology information LRR domain binding / microfibril binding / RNA Polymerase III Chain Elongation / MMXD complex / core TFIIH complex portion of holo TFIIH complex / RNA Polymerase III Transcription Termination / Cytosolic iron-sulfur cluster assembly / nucleotide-excision repair, DNA duplex unwinding / central nervous system myelin formation / regulation of transcription by RNA polymerase I ...LRR domain binding / microfibril binding / RNA Polymerase III Chain Elongation / MMXD complex / core TFIIH complex portion of holo TFIIH complex / RNA Polymerase III Transcription Termination / Cytosolic iron-sulfur cluster assembly / nucleotide-excision repair, DNA duplex unwinding / central nervous system myelin formation / regulation of transcription by RNA polymerase I / positive regulation of core promoter binding / RNA polymerase II core complex assembly / positive regulation of mitotic recombination / meiotic sister chromatid cohesion / hair follicle maturation / RNA polymerase transcription factor SL1 complex / hair cell differentiation / RPAP3/R2TP/prefoldin-like complex / nucleotide-excision repair factor 3 complex / transcription factor TFIIE complex / phosphatase activator activity / Cytosolic sensors of pathogen-associated DNA / nucleotide-excision repair, preincision complex assembly / CAK-ERCC2 complex / RNA polymerase III general transcription initiation factor activity / UV protection / embryonic cleavage / transcription open complex formation at RNA polymerase II promoter / RNA polymerase I core promoter sequence-specific DNA binding / TFIIF-class transcription factor complex binding / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / transcriptional start site selection at RNA polymerase II promoter / RNA Polymerase III Transcription Initiation From Type 3 Promoter / transcription factor TFIIF complex / DNA 5'-3' helicase / RNA Polymerase III Abortive And Retractive Initiation / transcription factor TFIIA complex / female germ cell nucleus / G protein-coupled receptor internalization / male pronucleus / female pronucleus / transcription factor TFIIH holo complex / transcription factor TFIIH core complex / Abortive elongation of HIV-1 transcript in the absence of Tat / RNA polymerase II general transcription initiation factor binding / germinal vesicle / DNA 3'-5' helicase / RNA Polymerase I Transcription Termination / transcription preinitiation complex / FGFR2 alternative splicing / nuclear thyroid hormone receptor binding / MicroRNA (miRNA) biogenesis / Signaling by FGFR2 IIIa TM / regulation of mitotic cell cycle phase transition / Viral Messenger RNA Synthesis / hematopoietic stem cell proliferation / 3'-5' DNA helicase activity / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / transcription factor TFIID complex / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / RNA polymerase II general transcription initiation factor activity / bone mineralization / cell division site / spinal cord development / mRNA Capping / protein acetylation / PIWI-interacting RNA (piRNA) biogenesis / 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 / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / erythrocyte maturation / mRNA Splicing - Minor Pathway / RNA polymerase II complex binding / acetyltransferase activity / RNA Polymerase I Transcription Initiation / viral transcription / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / transcription by RNA polymerase I / aryl hydrocarbon receptor binding / nuclear-transcribed mRNA catabolic process / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / ATPase activator activity / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / Processing of Capped Intron-Containing Pre-mRNA / DNA topological change / TFIIB-class transcription factor binding / transcription by RNA polymerase III / intrinsic apoptotic signaling pathway by p53 class mediator / RNA polymerase II transcribes snRNA genes / RNA polymerase II activity Similarity search - Function | |||||||||||||||
Biological species | Homo sapiens (human) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 6.3 Å | |||||||||||||||
Authors | He Y / Yan C | |||||||||||||||
Funding support | United States, 4 items
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Citation | Journal: Nature / Year: 2016 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
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_8134.map.gz | 1.3 MB | EMDB map data format | |
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Header (meta data) | emd-8134-v30.xml emd-8134.xml | 54.3 KB 54.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_8134_fsc.xml | 6.9 KB | Display | FSC data file |
Images | emd_8134.png | 151.5 KB | ||
Filedesc metadata | emd-8134.cif.gz | 14 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-8134 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8134 | HTTPS FTP |
-Validation report
Summary document | emd_8134_validation.pdf.gz | 380.1 KB | Display | EMDB validaton report |
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Full document | emd_8134_full_validation.pdf.gz | 379.6 KB | Display | |
Data in XML | emd_8134_validation.xml.gz | 9.2 KB | Display | |
Data in CIF | emd_8134_validation.cif.gz | 12 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8134 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8134 | HTTPS FTP |
-Related structure data
Related structure data | 5iy9MC 8131C 8132C 8133C 8135C 8136C 8137C 8138C 5ivwC 5iy6C 5iy7C 5iy8C 5iyaC 5iybC 5iycC 5iydC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_8134.map.gz / Format: CCP4 / Size: 27 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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
-Sample components
+Entire : Human holo-PIC in the initial transcribing state (no IIS)
+Supramolecule #1: Human holo-PIC in the initial transcribing state (no IIS)
+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: TFIIH basal transcription factor complex helicase XPB subunit
+Macromolecule #22: TFIIH basal transcription factor complex helicase XPD subunit
+Macromolecule #23: General transcription factor IIH subunit 2
+Macromolecule #24: General transcription factor IIH subunit 5
+Macromolecule #25: General transcription factor IIH subunit 4
+Macromolecule #26: General transcription factor IIH subunit 3
+Macromolecule #27: SCP-X
+Macromolecule #28: SCP-Y
+Macromolecule #29: RNA
+Macromolecule #30: MAGNESIUM ION
+Macromolecule #31: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-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).. |
-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: FIELD EMISSION GUN |
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 |
+Image processing
-Atomic model buiding 1
Refinement | Protocol: FLEXIBLE FIT |
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Output model | PDB-5iy9: |