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Yorodumi- EMDB-3376: Transcription initiation complex structures elucidate promoter DN... -
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Basic information
| Entry | Database: EMDB / ID: EMD-3376 | |||||||||
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| Title | Transcription initiation complex structures elucidate promoter DNA opening (OC2) | |||||||||
Map data | sharpened OC2 map | |||||||||
Sample |
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Keywords | Gene expression / Transcription initiation | |||||||||
| Function / homology | Function and homology informationTFIIA-class transcription factor complex binding / transcription open complex formation at RNA polymerase II promoter / RNA polymerase III transcription regulatory region sequence-specific DNA binding / RNA polymerase III preinitiation complex assembly / RNA polymerase I general transcription initiation factor binding / transcription factor TFIIE complex / TFIIF-class transcription factor complex binding / transcriptional start site selection at RNA polymerase II promoter / transcription factor TFIIF complex / transcription factor TFIIIB complex ...TFIIA-class transcription factor complex binding / transcription open complex formation at RNA polymerase II promoter / RNA polymerase III transcription regulatory region sequence-specific DNA binding / RNA polymerase III preinitiation complex assembly / RNA polymerase I general transcription initiation factor binding / transcription factor TFIIE complex / TFIIF-class transcription factor complex binding / transcriptional start site selection at RNA polymerase II promoter / transcription factor TFIIF complex / transcription factor TFIIIB complex / transcription factor TFIIA complex / RNA polymerase I preinitiation complex assembly / DNA binding, bending / transcription preinitiation complex / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / RNA Polymerase I Transcription Initiation / transcription factor TFIID complex / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA polymerase II general transcription initiation factor activity / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / mRNA Capping / Estrogen-dependent gene expression / Formation of TC-NER Pre-Incision Complex / RNA polymerase II transcribes snRNA genes / RNA Polymerase I Promoter Escape / TP53 Regulates Transcription of DNA Repair Genes / 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 / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / Gap-filling DNA repair synthesis and ligation in TC-NER / RNA polymerase II complex binding / termination of RNA polymerase II transcription / protein phosphatase activator activity / termination of RNA polymerase I transcription / Dual incision in TC-NER / positive regulation of transcription initiation by RNA polymerase II / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / positive regulation of translational initiation / nucleolar large rRNA transcription by RNA polymerase I / transcription initiation at RNA polymerase I promoter / transcription by RNA polymerase III / nuclear-transcribed mRNA catabolic process / RNA polymerase II core promoter sequence-specific DNA binding / RNA polymerase II preinitiation complex assembly / termination of RNA polymerase III transcription / transcription initiation at RNA polymerase III promoter / RNA polymerase I complex / RNA polymerase III complex / RNA polymerase II, core complex / transcription elongation by RNA polymerase I / tRNA transcription by RNA polymerase III / transcription by RNA polymerase I / transcription-coupled nucleotide-excision repair / translesion synthesis / translation initiation factor binding / TBP-class protein binding / DNA-templated transcription elongation / positive regulation of transcription elongation by RNA polymerase II / DNA-templated transcription initiation / P-body / transcription initiation at RNA polymerase II promoter / mRNA processing / transcription elongation by RNA polymerase II / mRNA transcription by RNA polymerase II / transcription by RNA polymerase II / ribonucleoside binding / transcription coregulator activity / disordered domain specific binding / cytoplasmic stress granule / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / peroxisome / single-stranded DNA binding / transcription regulator complex / ribosome biogenesis / DNA-binding transcription factor binding / RNA polymerase II-specific DNA-binding transcription factor binding / nucleic acid binding / transcription coactivator activity / protein dimerization activity / single-stranded RNA binding / nucleotide binding / negative regulation of DNA-templated transcription / mRNA binding / chromatin binding / nucleolus / regulation of DNA-templated transcription / positive regulation of transcription by RNA polymerase II / protein-containing complex / mitochondrion / DNA binding / metal ion binding / zinc ion binding Similarity search - Function | |||||||||
| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.97 Å | |||||||||
Authors | Plaschka C / Hantsche M / Dienemann C / Burzinski C / Plitzko J / Cramer P | |||||||||
Citation | Journal: Nature / Year: 2016Title: Transcription initiation complex structures elucidate DNA opening. Authors: C Plaschka / M Hantsche / C Dienemann / C Burzinski / J Plitzko / P Cramer / ![]() Abstract: Transcription of eukaryotic protein-coding genes begins with assembly of the RNA polymerase (Pol) II initiation complex and promoter DNA opening. Here we report cryo-electron microscopy (cryo-EM) ...Transcription of eukaryotic protein-coding genes begins with assembly of the RNA polymerase (Pol) II initiation complex and promoter DNA opening. Here we report cryo-electron microscopy (cryo-EM) structures of yeast initiation complexes containing closed and open DNA at resolutions of 8.8 Å and 3.6 Å, respectively. DNA is positioned and retained over the Pol II cleft by a network of interactions between the TATA-box-binding protein TBP and transcription factors TFIIA, TFIIB, TFIIE, and TFIIF. DNA opening occurs around the tip of the Pol II clamp and the TFIIE 'extended winged helix' domain, and can occur in the absence of TFIIH. Loading of the DNA template strand into the active centre may be facilitated by movements of obstructing protein elements triggered by allosteric binding of the TFIIE 'E-ribbon' domain. The results suggest a unified model for transcription initiation with a key event, the trapping of open promoter DNA by extended protein-protein and protein-DNA contacts. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_3376.map.gz | 96.4 MB | EMDB map data format | |
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| Header (meta data) | emd-3376-v30.xml emd-3376.xml | 14 KB 14 KB | Display Display | EMDB header |
| Images | emd_3376.png | 200.5 KB | ||
| Others | emd_3376_additional_1.map.gz | 95.3 MB | ||
| Archive directory | https://data.pdbj.org/pub/emdb/structures/EMD-3376 ftp://data.pdbj.org/pub/emdb/structures/EMD-3376 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 3375C ![]() 3377C ![]() 3378C ![]() 3379C ![]() 3380C ![]() 3381C ![]() 3382C ![]() 3383C ![]() 5fywC ![]() 5fz5C ![]() 5ip7C ![]() 5ip9C C: citing same article ( |
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| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_3376.map.gz / Format: CCP4 / Size: 100.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | sharpened OC2 map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.35 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Supplemental map: emd 3376 additional 1.map
| File | emd_3376_additional_1.map | ||||||||||||
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| Projections & Slices |
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| Density Histograms |
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Sample components
-Entire : yeast Pol II transcription initiation complex (open DNA)
| Entire | Name: yeast Pol II transcription initiation complex (open DNA) |
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| Components |
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-Supramolecule #1000: yeast Pol II transcription initiation complex (open DNA)
| Supramolecule | Name: yeast Pol II transcription initiation complex (open DNA) type: sample / ID: 1000 / Oligomeric state: Monomer / Number unique components: 7 |
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| Molecular weight | Theoretical: 890 KDa |
-Macromolecule #1: DNA-directed RNA polymerase II
| Macromolecule | Name: DNA-directed RNA polymerase II / type: protein_or_peptide / ID: 1 / Name.synonym: RNA polymerase II / Number of copies: 1 / Recombinant expression: No |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 513 KDa |
-Macromolecule #2: Transcription Factor IIA
| Macromolecule | Name: Transcription Factor IIA / type: protein_or_peptide / ID: 2 / Name.synonym: TFIIA / Number of copies: 1 / Recombinant expression: Yes |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 45 KDa |
| Recombinant expression | Organism: ![]() |
-Macromolecule #3: Transcription Factor IIB
| Macromolecule | Name: Transcription Factor IIB / type: protein_or_peptide / ID: 3 / Name.synonym: TFIIB / Number of copies: 1 / Recombinant expression: Yes |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 38 KDa |
| Recombinant expression | Organism: ![]() |
-Macromolecule #4: TATA-box-binding protein
| Macromolecule | Name: TATA-box-binding protein / type: protein_or_peptide / ID: 4 / Name.synonym: TBP, SPT15 / Number of copies: 1 / Recombinant expression: Yes |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 27 KDa |
| Recombinant expression | Organism: ![]() |
-Macromolecule #5: Transcription Factor IIE
| Macromolecule | Name: Transcription Factor IIE / type: protein_or_peptide / ID: 5 / Name.synonym: TFIIE / Number of copies: 1 / Recombinant expression: Yes |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 91 KDa |
| Recombinant expression | Organism: ![]() |
-Macromolecule #6: Transcription Factor IIF
| Macromolecule | Name: Transcription Factor IIF / type: protein_or_peptide / ID: 6 / Name.synonym: TFIIF / Number of copies: 1 / Recombinant expression: Yes |
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| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 129 KDa |
| Recombinant expression | Organism: ![]() |
-Macromolecule #7: synthetic promoter DNA construct (open)
| Macromolecule | Name: synthetic promoter DNA construct (open) / type: dna / ID: 7 / Details: Contains a 15 nucleotide mismatch bubble / Classification: DNA / Structure: DOUBLE HELIX / Synthetic?: Yes |
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| Source (natural) | Organism: unidentified (others) |
| Molecular weight | Theoretical: 44 KDa |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.3 mg/mL |
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| Buffer | pH: 7.5 Details: 25 mM HEPES-KOH pH 7.5, 150 mM potassium acetate, 2 mM MgCl2, 5 mM DTT |
| Grid | Details: R3.5/1 holey carbon grids (Quantifoil) |
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV Method: Quantifoil R 3.5/1 holey carbon grids were glow-discharged before deposition of 4.5 microliters of sample. Grids were then blotted for 8.5 s and plunge-frozen in liquid ethane. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Specialist optics | Energy filter - Name: GIF Quantum / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 20.0 eV |
| Date | Apr 26, 2016 |
| Image recording | Category: CCD / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Number real images: 1756 / Average electron dose: 33 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Calibrated magnification: 37037 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.3 mm / Nominal defocus max: 4.2 µm / Nominal defocus min: 0.7 µm / Nominal magnification: 37000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| CTF correction | Details: Each particle |
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| Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 3.97 Å / Resolution method: OTHER / Software - Name: RELION / Number images used: 17282 |
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