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Open data
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Basic information
| Entry | Database: PDB / ID: 9qcs | |||||||||||||||||||||||||||
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| Title | Ku80 mediated DNA-PK dimer bound to 153 bp H2AX nucleosome | |||||||||||||||||||||||||||
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Keywords | DNA BINDING PROTEIN / DNA-binding protein / NHEJ / Ku70/80 / nucleosome | |||||||||||||||||||||||||||
| Function / homology | Function and homology informationpositive regulation of platelet formation / Ku70:Ku80 complex / T cell receptor V(D)J recombination / negative regulation of t-circle formation / pro-B cell differentiation / DNA end binding / DNA-dependent protein kinase activity / small-subunit processome assembly / positive regulation of lymphocyte differentiation / histone H2AXS139 kinase activity ...positive regulation of platelet formation / Ku70:Ku80 complex / T cell receptor V(D)J recombination / negative regulation of t-circle formation / pro-B cell differentiation / DNA end binding / DNA-dependent protein kinase activity / small-subunit processome assembly / positive regulation of lymphocyte differentiation / histone H2AXS139 kinase activity / DNA-dependent protein kinase complex / DNA-dependent protein kinase-DNA ligase 4 complex / immunoglobulin V(D)J recombination / nonhomologous end joining complex / immature B cell differentiation / regulation of smooth muscle cell proliferation / cellular response to X-ray / regulation of epithelial cell proliferation / double-strand break repair via alternative nonhomologous end joining / XY body / double-strand break repair via classical nonhomologous end joining / nuclear telomere cap complex / protein localization to site of double-strand break / chromatin-protein adaptor activity / Cytosolic sensors of pathogen-associated DNA / telomere capping / IRF3-mediated induction of type I IFN / regulation of hematopoietic stem cell differentiation / recombinational repair / positive regulation of neurogenesis / regulation of telomere maintenance / U3 snoRNA binding / protein localization to chromosome, telomeric region / maturation of 5.8S rRNA / T cell lineage commitment / cellular hyperosmotic salinity response / negative regulation of cGAS/STING signaling pathway / positive regulation of double-strand break repair via nonhomologous end joining / response to ionizing radiation / B cell lineage commitment / 2-LTR circle formation / hematopoietic stem cell proliferation / telomeric DNA binding / negative regulation of protein phosphorylation / positive regulation of protein kinase activity / Lyases; Carbon-oxygen lyases; Other carbon-oxygen lyases / site of DNA damage / peptidyl-threonine phosphorylation / 5'-deoxyribose-5-phosphate lyase activity / hematopoietic stem cell differentiation / negative regulation of tumor necrosis factor-mediated signaling pathway / ATP-dependent activity, acting on DNA / somitogenesis / ectopic germ cell programmed cell death / telomere maintenance via telomerase / negative regulation of megakaryocyte differentiation / protein localization to CENP-A containing chromatin / Chromatin modifying enzymes / Replacement of protamines by nucleosomes in the male pronucleus / CENP-A containing nucleosome / mitotic G1 DNA damage checkpoint signaling / Packaging Of Telomere Ends / neurogenesis / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / Deposition of new CENPA-containing nucleosomes at the centromere / telomere organization / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / Interleukin-7 signaling / activation of innate immune response / DNA helicase activity / telomere maintenance / RNA Polymerase I Promoter Opening / epigenetic regulation of gene expression / positive regulation of erythrocyte differentiation / Inhibition of DNA recombination at telomere / Assembly of the ORC complex at the origin of replication / cyclin binding / Meiotic synapsis / DNA damage checkpoint signaling / SUMOylation of chromatin organization proteins / Regulation of endogenous retroelements by the Human Silencing Hub (HUSH) complex / positive regulation of DNA repair / DNA methylation / replication fork / positive regulation of translation / Condensation of Prophase Chromosomes / cellular response to leukemia inhibitory factor / Chromatin modifications during the maternal to zygotic transition (MZT) / SIRT1 negatively regulates rRNA expression / HCMV Late Events / condensed nuclear chromosome / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / response to gamma radiation / PRC2 methylates histones and DNA / Regulation of endogenous retroelements by KRAB-ZFP proteins / innate immune response in mucosa / Defective pyroptosis / meiotic cell cycle Similarity search - Function | |||||||||||||||||||||||||||
| Biological species | Homo sapiens (human) | |||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.38 Å | |||||||||||||||||||||||||||
Authors | Hall, C. / Chaplin, A.K. | |||||||||||||||||||||||||||
| Funding support | Canada, United Kingdom, 2items
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Citation | Journal: Nat Commun / Year: 2025Title: Cryo-EM structures of NHEJ assemblies with nucleosomes. Authors: Chloe Hall / Philippe Frit / Antonia Kefala-Stavridi / Amandine Pelletier / Steven W Hardwick / Himani Amin / Matthew K Bilyard / Taiana Maia De Oliviera / Ammarah Tariq / Sayma Zahid / ...Authors: Chloe Hall / Philippe Frit / Antonia Kefala-Stavridi / Amandine Pelletier / Steven W Hardwick / Himani Amin / Matthew K Bilyard / Taiana Maia De Oliviera / Ammarah Tariq / Sayma Zahid / Dimitri Y Chirgadze / Shankar Balasubramanian / Katheryn Meek / Virginie Ropars / Jean-Baptiste Charbonnier / Mauro Modesti / Patrick Calsou / Sébastien Britton / Tom L Blundell / Thomas Schalch / Amanda K Chaplin / ![]() Abstract: DNA double-strand breaks (DSBs) are highly deleterious lesions that can trigger cell death or carcinogenesis if unrepaired or misrepaired. In mammals, most DSBs are repaired by non-homologous end ...DNA double-strand breaks (DSBs) are highly deleterious lesions that can trigger cell death or carcinogenesis if unrepaired or misrepaired. In mammals, most DSBs are repaired by non-homologous end joining (NHEJ), which begins when Ku70/80 binds DNA ends and recruits DNA-PKcs to form the DNA-PK holoenzyme. Although recent cryo-EM studies have resolved several NHEJ assemblies, how these factors access DSBs within nucleosomes remains unclear. Here, we present cryo-EM structures of human Ku70/80 and DNA-PK bound to nucleosomes. Ku70/80 binds the DNA end and bends it away from the nucleosome core, while the Ku70 C-terminal SAP domain makes an additional, specific DNA contact. Our DNA-PK-nucleosome structure further reveals the opening of the Ku80 vWA domain, and we show that non-hydrolysable ATP promotes synapsis by stabilising the Ku80-mediated DNA-PK dimer. These structures reveal a model for DSB recognition on nucleosomal DNA and provide insights relevant to targeting NHEJ in cancer therapy. | |||||||||||||||||||||||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 9qcs.cif.gz | 2 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb9qcs.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 9qcs.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 9qcs_validation.pdf.gz | 1.7 MB | Display | wwPDB validaton report |
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| Full document | 9qcs_full_validation.pdf.gz | 2.1 MB | Display | |
| Data in XML | 9qcs_validation.xml.gz | 301.1 KB | Display | |
| Data in CIF | 9qcs_validation.cif.gz | 473.9 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/qc/9qcs ftp://data.pdbj.org/pub/pdb/validation_reports/qc/9qcs | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 53026MC ![]() 9igwC ![]() 9igxC ![]() 9q80C ![]() 9q8xC ![]() 9q9fC ![]() 9qcrC ![]() 9qmsC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-Protein , 5 types, 18 molecules OoDHdhCGcgAEaeBFbf
| #1: Protein | Mass: 469673.219 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)References: UniProt: P78527, non-specific serine/threonine protein kinase #2: Protein | Mass: 13935.239 Da / Num. of mol.: 4 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: H2BC11, H2BFR, HIST1H2BJ / Production host: ![]() #3: Protein | Mass: 15172.605 Da / Num. of mol.: 4 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: H2AX, H2AFX / Production host: ![]() #4: Protein | Mass: 15437.167 Da / Num. of mol.: 4 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human)Gene: HIST1H3A, H3FA, HIST1H3B, H3FL, HIST1H3C, H3FC, HIST1H3D, H3FB, HIST1H3E, H3FD, HIST1H3F, H3FI, HIST1H3G, H3FH, HIST1H3H, H3FK, HIST1H3I, H3FF, HIST1H3J, H3FJ Production host: ![]() #5: Protein | Mass: 11394.426 Da / Num. of mol.: 4 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human)Gene: H4C1, H4/A, H4FA, HIST1H4A, H4C2, H4/I, H4FI, HIST1H4B, H4C3, H4/G, H4FG, HIST1H4C, H4C4, H4/B, H4FB, HIST1H4D, H4C5, H4/J, H4FJ, HIST1H4E, H4C6, H4/C, H4FC, HIST1H4F, H4C8, H4/H, H4FH, ...Gene: H4C1, H4/A, H4FA, HIST1H4A, H4C2, H4/I, H4FI, HIST1H4B, H4C3, H4/G, H4FG, HIST1H4C, H4C4, H4/B, H4FB, HIST1H4D, H4C5, H4/J, H4FJ, HIST1H4E, H4C6, H4/C, H4FC, HIST1H4F, H4C8, H4/H, H4FH, HIST1H4H, H4C9, H4/M, H4FM, HIST1H4I, H4C11, H4/E, H4FE, HIST1H4J, H4C12, H4/D, H4FD, HIST1H4K, H4C13, H4/K, H4FK, HIST1H4L, H4C14, H4/N, H4F2, H4FN, HIST2H4, HIST2H4A, H4C15, H4/O, H4FO, HIST2H4B, H4C16, H4-16, HIST4H4 Production host: ![]() |
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-DNA chain , 2 types, 4 molecules iIjJ
| #6: DNA chain | Mass: 47457.234 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Production host: ![]() #7: DNA chain | Mass: 46998.945 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Production host: ![]() |
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-X-ray repair cross-complementing protein ... , 2 types, 4 molecules KMLN
| #8: Protein | Mass: 69945.039 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: XRCC6, G22P1 / Production host: ![]() References: UniProt: P12956, Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement, Lyases; Carbon-oxygen lyases; Other carbon-oxygen lyases #9: Protein | Mass: 82812.438 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: XRCC5, G22P2 / Production host: ![]() References: UniProt: P13010, Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement |
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-Details
| Has protein modification | N |
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-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: DNA-PK dimer bound to 153 bp H2AX nucleosome / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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| Molecular weight | Value: 1.640 MDa / Experimental value: YES |
| Source (natural) | Organism: Homo sapiens (human) |
| Source (recombinant) | Organism: ![]() |
| Buffer solution | pH: 7.6 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Specimen support | Details: 30mA / Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: TFS KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2200 nm / Nominal defocus min: 700 nm |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Electron dose: 48.59 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 | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 4.38 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 27712 / Symmetry type: POINT | ||||||||||||||||||||||||
| Refinement | Highest resolution: 4.38 Å Stereochemistry target values: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) | ||||||||||||||||||||||||
| Refine LS restraints |
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About Yorodumi




Homo sapiens (human)
Canada,
United Kingdom, 2items
Citation
















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FIELD EMISSION GUN