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Open data
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Basic information
| Entry | Database: PDB / ID: 7k04 | ||||||
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| Title | Structure of TFIIH/Rad4-Rad23-Rad33/DNA in DNA opening | ||||||
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Keywords | NUCLEAR PROTEIN/DNA / TFIIH / Rad4 / Rad4/23 / XPC / NER / Nucleotide Excision Repair / GG-NER / NUCLEAR PROTEIN / NUCLEAR PROTEIN-DNA complex | ||||||
| Function / homology | Function and homology informationnucleotide-excision repair factor 2 complex / single-strand break-containing DNA binding / XPC complex / nucleotide-excision repair, DNA damage recognition / regulation of mitotic recombination / transcription open complex formation at RNA polymerase II promoter / phosphatidylinositol-5-phosphate binding / RNA polymerase II promoter clearance / positive regulation of mitotic recombination / nucleotide-excision repair factor 3 complex ...nucleotide-excision repair factor 2 complex / single-strand break-containing DNA binding / XPC complex / nucleotide-excision repair, DNA damage recognition / regulation of mitotic recombination / transcription open complex formation at RNA polymerase II promoter / phosphatidylinositol-5-phosphate binding / RNA polymerase II promoter clearance / positive regulation of mitotic recombination / nucleotide-excision repair factor 3 complex / nucleotide-excision repair, preincision complex assembly / DNA translocase activity / transcriptional start site selection at RNA polymerase II promoter / DNA 5'-3' helicase / phosphatidylinositol-3-phosphate binding / transcription factor TFIIH core complex / transcription factor TFIIH holo complex / transcription preinitiation complex / poly(A)+ mRNA export from nucleus / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / mRNA Capping / DNA 3'-5' helicase / 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 / 3'-5' DNA helicase activity / Formation of TC-NER Pre-Incision Complex / RNA Polymerase I Promoter Escape / Gap-filling DNA repair synthesis and ligation in TC-NER / ATPase activator activity / DNA topological change / Dual incision in TC-NER / ATP-dependent activity, acting on DNA / mismatch repair / transcription by RNA polymerase I / DNA helicase activity / nucleotide-excision repair / transcription initiation at RNA polymerase II promoter / ubiquitin protein ligase activity / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / double-stranded DNA binding / 5'-3' DNA helicase activity / proteasome-mediated ubiquitin-dependent protein catabolic process / damaged DNA binding / transcription by RNA polymerase II / DNA repair / regulation of DNA-templated transcription / regulation of transcription by RNA polymerase II / negative regulation of transcription by RNA polymerase II / ATP hydrolysis activity / DNA binding / zinc ion binding / ATP binding / metal ion binding / nucleus / cytoplasm / cytosol Similarity search - Function | ||||||
| Biological species | ![]() synthetic construct (others) | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 9.25 Å | ||||||
Authors | van Eeuwen, T. / Min, J.H. / Murakami, K. | ||||||
| Funding support | United States, 1items
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Citation | Journal: Nat Commun / Year: 2021Title: Cryo-EM structure of TFIIH/Rad4-Rad23-Rad33 in damaged DNA opening in nucleotide excision repair. Authors: Trevor van Eeuwen / Yoonjung Shim / Hee Jong Kim / Tingting Zhao / Shrabani Basu / Benjamin A Garcia / Craig D Kaplan / Jung-Hyun Min / Kenji Murakami / ![]() Abstract: The versatile nucleotide excision repair (NER) pathway initiates as the XPC-RAD23B-CETN2 complex first recognizes DNA lesions from the genomic DNA and recruits the general transcription factor ...The versatile nucleotide excision repair (NER) pathway initiates as the XPC-RAD23B-CETN2 complex first recognizes DNA lesions from the genomic DNA and recruits the general transcription factor complex, TFIIH, for subsequent lesion verification. Here, we present a cryo-EM structure of an NER initiation complex containing Rad4-Rad23-Rad33 (yeast homologue of XPC-RAD23B-CETN2) and 7-subunit coreTFIIH assembled on a carcinogen-DNA adduct lesion at 3.9-9.2 Å resolution. A ~30-bp DNA duplex could be mapped as it straddles between Rad4 and the Ssl2 (XPB) subunit of TFIIH on the 3' and 5' side of the lesion, respectively. The simultaneous binding with Rad4 and TFIIH was permitted by an unwinding of DNA at the lesion. Translocation coupled with torque generation by Ssl2 and Rad4 would extend the DNA unwinding at the lesion and deliver the damaged strand to Rad3 (XPD) in an open form suitable for subsequent lesion scanning and verification. | ||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7k04.cif.gz | 787.7 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb7k04.ent.gz | 608 KB | Display | PDB format |
| PDBx/mmJSON format | 7k04.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 7k04_validation.pdf.gz | 1.5 MB | Display | wwPDB validaton report |
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| Full document | 7k04_full_validation.pdf.gz | 1.6 MB | Display | |
| Data in XML | 7k04_validation.xml.gz | 120.5 KB | Display | |
| Data in CIF | 7k04_validation.cif.gz | 184.3 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/k0/7k04 ftp://data.pdbj.org/pub/pdb/validation_reports/k0/7k04 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 22588MC ![]() 7k01C ![]() 7m2uC C: citing same article ( M: map data used to model this data |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-DNA repair protein ... , 2 types, 2 molecules EA
| #1: Protein | Mass: 20291.457 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Strain: ATCC 204508 / S288c / Gene: RAD33, YML011C, YM9571.07C / Production host: ![]() |
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| #9: Protein | Mass: 87374.492 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Strain: ATCC 204508 / S288c / Gene: RAD4, YER162C / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: P14736 |
-General transcription and DNA repair factor IIH subunit ... , 4 types, 4 molecules 2146
| #2: Protein | Mass: 58602.312 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) ![]() Strain: ATCC 204508 / S288c / References: UniProt: Q02939 |
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| #4: Protein | Mass: 72959.258 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) ![]() Strain: ATCC 204508 / S288c / References: UniProt: P32776 |
| #5: Protein | Mass: 37506.422 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) ![]() Strain: ATCC 204508 / S288c / References: UniProt: Q12004 |
| #6: Protein | Mass: 52370.035 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) ![]() Strain: ATCC 204508 / S288c / References: UniProt: Q04673 |
-DNA repair helicase ... , 2 types, 2 molecules 07
| #3: Protein | Mass: 89899.047 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) ![]() Strain: ATCC 204508 / S288c / References: UniProt: P06839, DNA helicase |
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| #10: Protein | Mass: 95461.664 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Strain: ATCC 204508 / S288c / Gene: SSL2, LOM3, RAD25, UVS112, YIL143C / Production host: ![]() |
-DNA chain , 2 types, 2 molecules YW
| #7: DNA chain | Mass: 8641.573 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
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| #8: DNA chain | Mass: 8855.751 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
-Protein , 1 types, 1 molecules 5
| #11: Protein | Mass: 8243.490 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Strain: ATCC 204508 / S288c / Gene: TFB5, YDR079C-A / Production host: ![]() |
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-Non-polymers , 3 types, 8 molecules 




| #12: Chemical | | #13: Chemical | ChemComp-SF4 / | #14: Chemical | ChemComp-ZN / |
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-Details
| Has ligand of interest | N |
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| Has protein modification | N |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: 10 component complex of Rad4-23, Rad33 and TFIIH on damaged DNA Type: COMPLEX / Entity ID: #1-#10 / Source: MULTIPLE SOURCES |
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| Molecular weight | Experimental value: NO |
| Buffer solution | pH: 7.6 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Specimen support | Grid material: COPPER / Grid type: Quantifoil R1.2/1.3 |
| Vitrification | Instrument: LEICA EM CPC / Cryogen name: ETHANE / Details: Manually blotted by Leica EM CPC |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TITAN KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: SPOT SCAN |
| Electron lens | Mode: BRIGHT FIELD |
| Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 9.25 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 73146 / Symmetry type: POINT | ||||||||||||||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT | ||||||||||||||||||||||||
| Atomic model building | PDB-ID: 6CFI Accession code: 6CFI / Source name: PDB / Type: experimental model |
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Trichoplusia ni (cabbage looper)

