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- EMDB-12745: Yeast RNA polymerase II transcription pre-initiation complex with... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-12745 | ||||||||||||||||||
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Title | Yeast RNA polymerase II transcription pre-initiation complex with open promoter DNA | ||||||||||||||||||
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![]() | Pre-initiation complex / TRANSCRIPTION | ||||||||||||||||||
Function / homology | ![]() regulation of mitotic recombination / RNA polymerase II promoter clearance / RNA polymerase II complex recruiting activity / transcription open complex formation at RNA polymerase II promoter / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / phosphatidylinositol-5-phosphate binding / RNA polymerase III preinitiation complex assembly / transcription factor TFIIIB complex / positive regulation of mitotic recombination ...regulation of mitotic recombination / RNA polymerase II promoter clearance / RNA polymerase II complex recruiting activity / transcription open complex formation at RNA polymerase II promoter / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / phosphatidylinositol-5-phosphate binding / RNA polymerase III preinitiation complex assembly / transcription factor TFIIIB complex / positive regulation of mitotic recombination / nucleotide-excision repair factor 3 complex / RNA polymerase I general transcription initiation factor binding / transcription factor TFIIE complex / nucleotide-excision repair, preincision complex assembly / DNA translocase activity / regulation of transcription by RNA polymerase III / transcription factor TFIIK complex / TFIIF-class transcription factor complex binding / transcriptional start site selection at RNA polymerase II promoter / RPB4-RPB7 complex / transcription factor TFIIF complex / DNA 5'-3' helicase / phosphatidylinositol-3-phosphate binding / RNA polymerase I preinitiation complex assembly / transcription factor TFIIA complex / transcription factor TFIIH core complex / transcription factor TFIIH holo complex / cyclin-dependent protein serine/threonine kinase activator activity / : / DNA binding, bending / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / RNA Polymerase I Transcription Initiation / transcription preinitiation complex / 3'-5' DNA helicase activity / Processing of Capped Intron-Containing Pre-mRNA / DNA 3'-5' helicase / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / poly(A)+ mRNA export from nucleus / mRNA Capping / RNA polymerase II transcribes snRNA genes / TP53 Regulates Transcription of DNA Repair Genes / termination of RNA polymerase II transcription / 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 / RNA Polymerase II Pre-transcription Events / RNA-templated transcription / transcription factor TFIID complex / RNA polymerase II general transcription initiation factor activity / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / Formation of TC-NER Pre-Incision Complex / termination of RNA polymerase III transcription / RNA Polymerase I Promoter Escape / termination of RNA polymerase I transcription / transcription initiation at RNA polymerase III promoter / nucleolar large rRNA transcription by RNA polymerase I / Gap-filling DNA repair synthesis and ligation in TC-NER / RNA polymerase II complex binding / transcription initiation at RNA polymerase I promoter / Estrogen-dependent gene expression / ATPase activator activity / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / transcription by RNA polymerase I / nuclear-transcribed mRNA catabolic process / protein phosphatase activator activity / positive regulation of translational initiation / transcription by RNA polymerase III / Dual incision in TC-NER / ATP-dependent activity, acting on DNA / positive regulation of transcription initiation by RNA polymerase II / RNA polymerase I complex / tRNA transcription by RNA polymerase III / translesion synthesis / RNA polymerase III complex / transcription elongation by RNA polymerase I / RNA polymerase II core promoter sequence-specific DNA binding / transcription-coupled nucleotide-excision repair / DNA helicase activity / RNA polymerase II, core complex / : / RNA polymerase II preinitiation complex assembly / translation initiation factor binding / TBP-class protein binding / DNA-directed RNA polymerase activity / isomerase activity / nucleotide-excision repair / positive regulation of transcription elongation by RNA polymerase II / transcription initiation at RNA polymerase II promoter / DNA-templated transcription initiation / transcription coregulator activity / transcription elongation by RNA polymerase II / P-body / ribonucleoside binding / : / : / : / : Similarity search - Function | ||||||||||||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() ![]() | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||||||||||||||
![]() | Schilbach S / Aibara S | ||||||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structure of RNA polymerase II pre-initiation complex at 2.9 Å defines initial DNA opening. Authors: Sandra Schilbach / Shintaro Aibara / Christian Dienemann / Frauke Grabbe / Patrick Cramer / ![]() Abstract: Transcription initiation requires assembly of the RNA polymerase II (Pol II) pre-initiation complex (PIC) and opening of promoter DNA. Here, we present the long-sought high-resolution structure of ...Transcription initiation requires assembly of the RNA polymerase II (Pol II) pre-initiation complex (PIC) and opening of promoter DNA. Here, we present the long-sought high-resolution structure of the yeast PIC and define the mechanism of initial DNA opening. We trap the PIC in an intermediate state that contains half a turn of open DNA located 30-35 base pairs downstream of the TATA box. The initially opened DNA region is flanked and stabilized by the polymerase "clamp head loop" and the TFIIF "charged region" that both contribute to promoter-initiated transcription. TFIIE facilitates initiation by buttressing the clamp head loop and by regulating the TFIIH translocase. The initial DNA bubble is then extended in the upstream direction, leading to the open promoter complex and enabling start-site scanning and RNA synthesis. This unique mechanism of DNA opening may permit more intricate regulation than in the Pol I and Pol III systems. | ||||||||||||||||||
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
-Validation report
Summary document | ![]() | 708.1 KB | Display | ![]() |
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Full document | ![]() | 707.7 KB | Display | |
Data in XML | ![]() | 20.7 KB | Display | |
Data in CIF | ![]() | 26.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7o75MC ![]() 7o4iC ![]() 7o4jC ![]() 7o4kC ![]() 7o4lC ![]() 7o72C ![]() 7o73C 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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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.05 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
+Mask #1
+Additional map: Locally filtered map (non-composite) of an OC refinement...
+Additional map: Half-map 1 of focused refinement encompassing the Pol...
+Additional map: Half-map 2 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the Rad3/Ssl1...
+Additional map: Half-map 2 of focused refinement encompassing the Rad3/Ssl1...
+Additional map: Half-map 2 of focused refinement encompassing the Ssl1/Tfb4/Tfb1-3-helix-bundle...
+Additional map: Half-map 1 of focused refinement encompassing the Ssl1/Tfb4/Tfb1-3-helix-bundle...
+Additional map: Half-map 1 of focused refinement encompassing the Ssl2/Tfb2/Tfb5...
+Additional map: Half-map 2 of focused refinement encompassing the Ssl2/Tfb2/Tfb5...
+Additional map: Half-map 2 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the Pol...
+Additional map: Half-map 2 of focused refinement encompassing the TFIIE/Tfb1-PHD...
+Additional map: Half-map 1 of focused refinement encompassing the Tfb2...
+Additional map: Half-map 2 of focused refinement encompassing the Tfb2...
+Additional map: Half-map 2 of focused refinement encompassing the upstream...
+Additional map: Half-map 2 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the upstream...
+Additional map: Half-map 1 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the TFIIE/Tfb1-PHD...
+Additional map: Half-map 2 of focused refinement encompassing the Pol...
+Additional map: Half-map 2 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the Pol...
+Additional map: Half-map 2 of focused refinement encompassing the Pol...
+Additional map: Half-map 1 of focused refinement encompassing the Pol...
+Half map: #2
+Half map: #1
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Sample components
+Entire : Yeast RNA polymerase II transcription pre-initiation complex with...
+Supramolecule #1: Yeast RNA polymerase II transcription pre-initiation complex with...
+Macromolecule #1: General transcription and DNA repair factor IIH helicase subunit XPD
+Macromolecule #2: General transcription and DNA repair factor IIH subunit TFB1
+Macromolecule #3: General transcription and DNA repair factor IIH subunit TFB2
+Macromolecule #4: RNA polymerase II transcription factor B subunit 3
+Macromolecule #5: General transcription and DNA repair factor IIH subunit TFB4
+Macromolecule #6: General transcription and DNA repair factor IIH subunit TFB5
+Macromolecule #7: General transcription and DNA repair factor IIH subunit SSL1
+Macromolecule #8: General transcription and DNA repair factor IIH helicase subunit XPB
+Macromolecule #9: DNA-directed RNA polymerase II subunit RPB1
+Macromolecule #10: DNA-directed RNA polymerase II subunit RPB2
+Macromolecule #11: DNA-directed RNA polymerase II subunit RPB3
+Macromolecule #12: DNA-directed RNA polymerase II subunit RPB4
+Macromolecule #13: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+Macromolecule #14: DNA-directed RNA polymerases I, II, and III subunit RPABC2
+Macromolecule #15: DNA-directed RNA polymerase II subunit RPB7
+Macromolecule #16: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+Macromolecule #17: DNA-directed RNA polymerase II subunit RPB9
+Macromolecule #18: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+Macromolecule #19: DNA-directed RNA polymerase II subunit RPB11
+Macromolecule #20: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+Macromolecule #21: Transcription initiation factor IIB
+Macromolecule #23: TATA-box-binding protein
+Macromolecule #24: Transcription initiation factor IIF subunit alpha
+Macromolecule #25: Transcription initiation factor IIF subunit beta
+Macromolecule #27: Transcription initiation factor IIA large subunit
+Macromolecule #28: Transcription initiation factor IIA subunit 2
+Macromolecule #29: Transcription initiation factor IIE subunit alpha
+Macromolecule #30: Transcription initiation factor IIE subunit beta
+Macromolecule #22: Non-template DNA
+Macromolecule #26: Template DNA
+Macromolecule #31: IRON/SULFUR CLUSTER
+Macromolecule #32: ZINC ION
+Macromolecule #33: BERYLLIUM TRIFLUORIDE ION
+Macromolecule #34: MAGNESIUM ION
+Macromolecule #35: ADENOSINE-5'-DIPHOSPHATE
-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: 7.6 |
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Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 29670 / Average exposure time: 9.0 sec. / Average electron dose: 43.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 130000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |