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Yorodumi- EMDB-47262: Cryo-EM reconstruction of TC-NER transcription elongation complex... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-47262 | ||||||||||||
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Title | Cryo-EM reconstruction of TC-NER transcription elongation complex with STK19 (map III) | ||||||||||||
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Sample |
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Keywords | DNA repair / RNA polymerase II / Complex / Co-transcriptional process / TRANSCRIPTION | ||||||||||||
Biological species | Homo sapiens (human) / Sus scrofa (pig) / synthetic construct (others) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.3 Å | ||||||||||||
Authors | Mevissen TET / Kuemmecke M / Farnung L / Walter JC | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: Cell / Year: 2024 Title: STK19 positions TFIIH for cell-free transcription-coupled DNA repair. Authors: Tycho E T Mevissen / Maximilian Kümmecke / Ernst W Schmid / Lucas Farnung / Johannes C Walter / Abstract: In transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4, which cooperate with UVSSA and ELOF1 to recruit TFIIH. To explore the ...In transcription-coupled nucleotide excision repair (TC-NER), stalled RNA polymerase II (RNA Pol II) binds CSB and CRL4, which cooperate with UVSSA and ELOF1 to recruit TFIIH. To explore the mechanism of TC-NER, we recapitulated this reaction in vitro. When a plasmid containing a site-specific lesion is transcribed in frog egg extract, error-free repair is observed that depends on CSB, CRL4, UVSSA, and ELOF1. Repair also requires STK19, a factor previously implicated in transcription recovery after UV exposure. A 1.9-Å cryo-electron microscopy structure shows that STK19 binds the TC-NER complex through CSA and the RPB1 subunit of RNA Pol II. Furthermore, AlphaFold predicts that STK19 interacts with the XPD subunit of TFIIH, and disrupting this interface impairs cell-free repair. Molecular modeling suggests that STK19 positions TFIIH ahead of RNA Pol II for lesion verification. Our analysis of cell-free TC-NER suggests that STK19 couples RNA Pol II stalling to downstream repair events. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_47262.map.gz | 197.3 MB | EMDB map data format | |
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Header (meta data) | emd-47262-v30.xml emd-47262.xml | 16.2 KB 16.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_47262_fsc.xml | 15.3 KB | Display | FSC data file |
Images | emd_47262.png | 95.7 KB | ||
Masks | emd_47262_msk_1.map emd_47262_msk_2.map | 391 MB 391 MB | Mask map | |
Filedesc metadata | emd-47262.cif.gz | 4.3 KB | ||
Others | emd_47262_additional_1.map.gz emd_47262_half_map_1.map.gz emd_47262_half_map_2.map.gz | 369.1 MB 362.4 MB 362.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-47262 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-47262 | HTTPS FTP |
-Validation report
Summary document | emd_47262_validation.pdf.gz | 959 KB | Display | EMDB validaton report |
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Full document | emd_47262_full_validation.pdf.gz | 958.6 KB | Display | |
Data in XML | emd_47262_validation.xml.gz | 24.2 KB | Display | |
Data in CIF | emd_47262_validation.cif.gz | 31.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-47262 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-47262 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_47262.map.gz / Format: CCP4 / Size: 391 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.94 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_47262_msk_1.map | ||||||||||||
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-Mask #2
File | emd_47262_msk_2.map | ||||||||||||
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-Additional map: #1
File | emd_47262_additional_1.map | ||||||||||||
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-Half map: #1
File | emd_47262_half_map_1.map | ||||||||||||
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-Half map: #2
File | emd_47262_half_map_2.map | ||||||||||||
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-Sample components
-Entire : Transcription-coupled nucleotide excision repair complex
Entire | Name: Transcription-coupled nucleotide excision repair complex |
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Components |
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-Supramolecule #1: Transcription-coupled nucleotide excision repair complex
Supramolecule | Name: Transcription-coupled nucleotide excision repair complex type: complex / ID: 1 / Parent: 0 |
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-Supramolecule #2: CSA, DDB1
Supramolecule | Name: CSA, DDB1 / type: complex / ID: 2 / Parent: 1 |
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Source (natural) | Organism: Homo sapiens (human) |
-Supramolecule #3: CSB, UVSSA
Supramolecule | Name: CSB, UVSSA / type: complex / ID: 3 / Parent: 1 |
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Source (natural) | Organism: Homo sapiens (human) |
-Supramolecule #4: DDA1, ELOF1, STK19
Supramolecule | Name: DDA1, ELOF1, STK19 / type: complex / ID: 4 / Parent: 1 |
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Source (natural) | Organism: Homo sapiens (human) |
-Supramolecule #5: RNA Polymerase II proteins
Supramolecule | Name: RNA Polymerase II proteins / type: complex / ID: 5 / Parent: 1 |
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Source (natural) | Organism: Sus scrofa (pig) |
-Supramolecule #6: Nucleic Acid
Supramolecule | Name: Nucleic Acid / type: complex / ID: 6 / Parent: 1 |
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Source (natural) | Organism: synthetic construct (others) / Synthetically produced: Yes |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | TFS KRIOS |
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Specialist optics | Energy filter - Name: TFS Selectris / Energy filter - Slit width: 10 eV |
Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 52.4 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.6 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |