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- EMDB-0031: Structure of activated transcription complex Pol II-DSIF-PAF-SPT6... -
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Basic information
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Title | Structure of activated transcription complex Pol II-DSIF-PAF-SPT6 (Map A) | |||||||||||||||
![]() | Postprocessed map of overall refinement with an applied B factor of -98.65(Map A). | |||||||||||||||
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![]() | DNA / RNA Polymerase / DSIF / PAF1c / SPT6 / TRANSCRIPTION | |||||||||||||||
Function / homology | ![]() blastocyst growth / inner cell mass cell differentiation / Ski complex / RNA polymerase II C-terminal domain phosphoserine binding / mRNA decay by 3' to 5' exoribonuclease / negative regulation of DNA-templated transcription, elongation / regulation of isotype switching / Cdc73/Paf1 complex / regulation of muscle cell differentiation / nuclear-transcribed mRNA catabolic process, 3'-5' exonucleolytic nonsense-mediated decay ...blastocyst growth / inner cell mass cell differentiation / Ski complex / RNA polymerase II C-terminal domain phosphoserine binding / mRNA decay by 3' to 5' exoribonuclease / negative regulation of DNA-templated transcription, elongation / regulation of isotype switching / Cdc73/Paf1 complex / regulation of muscle cell differentiation / nuclear-transcribed mRNA catabolic process, 3'-5' exonucleolytic nonsense-mediated decay / regulation of mRNA export from nucleus / endodermal cell fate commitment / : / negative regulation of myeloid cell differentiation / DSIF complex / positive regulation of cell cycle G1/S phase transition / trophectodermal cell differentiation / blastocyst hatching / regulation of transcription elongation by RNA polymerase II / regulation of mRNA processing / nucleosome organization / Formation of RNA Pol II elongation complex / Formation of the Early Elongation Complex / Transcriptional regulation by small RNAs / RNA Polymerase II Pre-transcription Events / TP53 Regulates Transcription of DNA Repair Genes / FGFR2 alternative splicing / RNA polymerase II transcribes snRNA genes / mRNA Capping / mRNA Splicing - Minor Pathway / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Elongation / RNA Polymerase II Transcription Initiation And Promoter Clearance / RNA Pol II CTD phosphorylation and interaction with CE / Estrogen-dependent gene expression / Formation of TC-NER Pre-Incision Complex / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / mRNA Splicing - Major Pathway / blastocyst formation / mRNA 3'-end processing / positive regulation of DNA-templated transcription, elongation / Abortive elongation of HIV-1 transcript in the absence of Tat / : / transcription elongation-coupled chromatin remodeling / stem cell population maintenance / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / interleukin-6-mediated signaling pathway / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / mRNA Capping / negative regulation of gene expression, epigenetic / organelle membrane / RNA polymerase II complex binding / negative regulation of transcription elongation by RNA polymerase II / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / positive regulation of macroautophagy / positive regulation of translational initiation / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / RNA polymerase II transcribes snRNA genes / protein localization to nucleus / mRNA transport / Tat-mediated elongation of the HIV-1 transcript / cell surface receptor signaling pathway via JAK-STAT / Formation of HIV-1 elongation complex containing HIV-1 Tat / transcription elongation by RNA polymerase I / tRNA transcription by RNA polymerase III / RNA polymerase I complex / RNA polymerase III complex / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase II, core complex / transcription-coupled nucleotide-excision repair / core promoter sequence-specific DNA binding / nucleosome binding / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / translation initiation factor binding / rescue of stalled ribosome / RNA Polymerase II Pre-transcription Events / SH2 domain binding / : / : / : / RNA splicing / transcription elongation factor complex / : / positive regulation of transcription elongation by RNA polymerase II / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / P-body / : Similarity search - Function | |||||||||||||||
Biological species | ![]() ![]() ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||||||||
![]() | Vos SM / Farnung L | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structure of activated transcription complex Pol II-DSIF-PAF-SPT6. Authors: Seychelle M Vos / Lucas Farnung / Marc Boehning / Christoph Wigge / Andreas Linden / Henning Urlaub / Patrick Cramer / ![]() Abstract: Gene regulation involves activation of RNA polymerase II (Pol II) that is paused and bound by the protein complexes DRB sensitivity-inducing factor (DSIF) and negative elongation factor (NELF). Here ...Gene regulation involves activation of RNA polymerase II (Pol II) that is paused and bound by the protein complexes DRB sensitivity-inducing factor (DSIF) and negative elongation factor (NELF). Here we show that formation of an activated Pol II elongation complex in vitro requires the kinase function of the positive transcription elongation factor b (P-TEFb) and the elongation factors PAF1 complex (PAF) and SPT6. The cryo-EM structure of an activated elongation complex of Sus scrofa Pol II and Homo sapiens DSIF, PAF and SPT6 was determined at 3.1 Å resolution and compared to the structure of the paused elongation complex formed by Pol II, DSIF and NELF. PAF displaces NELF from the Pol II funnel for pause release. P-TEFb phosphorylates the Pol II linker to the C-terminal domain. SPT6 binds to the phosphorylated C-terminal-domain linker and opens the RNA clamp formed by DSIF. These results provide the molecular basis for Pol II pause release and elongation activation. | |||||||||||||||
History |
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Structure visualization
Structure viewer | EM map: ![]() ![]() ![]() |
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Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 15.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 54.9 KB 54.9 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() ![]() | 12.8 KB 12.8 KB | Display Display | ![]() |
Images | ![]() | 78.8 KB | ||
Masks | ![]() | 178 MB | ![]() | |
Filedesc metadata | ![]() | 14.2 KB | ||
Others | ![]() ![]() ![]() | 140.4 MB 140.9 MB 140.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 767.4 KB | Display | ![]() |
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Full document | ![]() | 767 KB | Display | |
Data in XML | ![]() | 14.9 KB | Display | |
Data in CIF | ![]() | 17.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6gmhMC ![]() 0030C ![]() 0032C ![]() 0033C ![]() 0034C ![]() 0035C ![]() 0036C ![]() 0037C ![]() 6gmeC M: atomic model generated by this map C: citing same article ( |
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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
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Annotation | Postprocessed map of overall refinement with an applied B factor of -98.65(Map A). | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.049 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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Density Histograms |
-Additional map: Global refinement of all EC* particles.
File | emd_0031_additional.map | ||||||||||||
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Annotation | Global refinement of all EC* particles. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 1 of global refinement (Map A).
File | emd_0031_half_map_1.map | ||||||||||||
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Annotation | Half map 1 of global refinement (Map A). | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2 of global refinement (Map A).
File | emd_0031_half_map_2.map | ||||||||||||
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Annotation | Half map 2 of global refinement (Map A). | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
+Entire : RNA Polymerase II-DSIF-PAF-SPT6 elongation complex (EC*)
+Supramolecule #1: RNA Polymerase II-DSIF-PAF-SPT6 elongation complex (EC*)
+Supramolecule #2: RNA Polymerase II
+Supramolecule #3: associated proteins
+Supramolecule #4: Nucleic acids
+Macromolecule #1: RPB1
+Macromolecule #2: DNA-directed RNA polymerase subunit beta
+Macromolecule #3: RNA polymerase II subunit C
+Macromolecule #4: RNA polymerase II subunit D
+Macromolecule #5: RNA polymerase II subunit E
+Macromolecule #6: RNA polymerase II subunit F
+Macromolecule #7: RNA polymerase II subunit G
+Macromolecule #8: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+Macromolecule #9: DNA-directed RNA polymerase II subunit RPB9
+Macromolecule #10: RPB10
+Macromolecule #11: RPB11
+Macromolecule #12: RPB12
+Macromolecule #13: Transcription elongation factor SPT6,Transcription elongation fac...
+Macromolecule #16: CTR9,RNA polymerase-associated protein CTR9 homolog,RNA polymeras...
+Macromolecule #18: LEO1,LEO1,RNA polymerase-associated protein LEO1
+Macromolecule #19: PAF1,RNA polymerase II-associated factor 1 homolog,RNA polymerase...
+Macromolecule #20: WD repeat-containing protein 61
+Macromolecule #21: CDC73
+Macromolecule #22: Transcription elongation factor SPT4
+Macromolecule #23: Transcription elongation factor SPT5
+Macromolecule #14: Non-template DNA
+Macromolecule #17: Template DNA
+Macromolecule #15: RNA
+Macromolecule #24: ZINC ION
+Macromolecule #25: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil UltrAuFoil / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: 10s wait prior to blotting, blotting 8.5s. |
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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 exposure time: 10.0 sec. / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |