+Open data
-Basic information
Entry | Database: PDB / ID: 8bah | |||||||||||||||
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Title | Human Mre11-Nbs1 complex | |||||||||||||||
Components |
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Keywords | HYDROLASE / DNA repair / complex | |||||||||||||||
Function / homology | Function and homology information telomere maintenance via telomere trimming / chromosomal region / mitochondrial double-strand break repair via homologous recombination / telomeric 3' overhang formation / Mre11 complex / blastocyst growth / meiotic DNA double-strand break formation / negative regulation of telomere capping / Sensing of DNA Double Strand Breaks / BRCA1-C complex ...telomere maintenance via telomere trimming / chromosomal region / mitochondrial double-strand break repair via homologous recombination / telomeric 3' overhang formation / Mre11 complex / blastocyst growth / meiotic DNA double-strand break formation / negative regulation of telomere capping / Sensing of DNA Double Strand Breaks / BRCA1-C complex / regulation of mitotic recombination / t-circle formation / DNA double-strand break processing / single-stranded DNA endodeoxyribonuclease activity / homologous chromosome pairing at meiosis / DNA strand resection involved in replication fork processing / nuclear inclusion body / homologous recombination / 5'-3' exonuclease activity / nuclease activity / 3'-5'-DNA exonuclease activity / positive regulation of telomere maintenance / HDR through MMEJ (alt-NHEJ) / isotype switching / Cytosolic sensors of pathogen-associated DNA / Impaired BRCA2 binding to PALB2 / IRF3-mediated induction of type I IFN / reciprocal meiotic recombination / mitotic G2/M transition checkpoint / positive regulation of kinase activity / mitotic intra-S DNA damage checkpoint signaling / sister chromatid cohesion / Defective homologous recombination repair (HRR) due to BRCA1 loss of function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA1 binding function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA2/RAD51/RAD51C binding function / Homologous DNA Pairing and Strand Exchange / Resolution of D-loop Structures through Synthesis-Dependent Strand Annealing (SDSA) / regulation of DNA-templated DNA replication initiation / Resolution of D-loop Structures through Holliday Junction Intermediates / DNA duplex unwinding / HDR through Single Strand Annealing (SSA) / Impaired BRCA2 binding to RAD51 / mitotic G2 DNA damage checkpoint signaling / Presynaptic phase of homologous DNA pairing and strand exchange / neuromuscular process controlling balance / neuroblast proliferation / telomere maintenance via telomerase / DNA damage response, signal transduction by p53 class mediator / positive regulation of protein autophosphorylation / 3'-5' exonuclease activity / telomere maintenance / intrinsic apoptotic signaling pathway / DNA endonuclease activity / replication fork / meiotic cell cycle / DNA damage checkpoint signaling / Nonhomologous End-Joining (NHEJ) / double-strand break repair via homologous recombination / HDR through Homologous Recombination (HRR) / G2/M DNA damage checkpoint / DNA Damage/Telomere Stress Induced Senescence / PML body / Meiotic recombination / double-strand break repair via nonhomologous end joining / double-strand break repair / site of double-strand break / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / manganese ion binding / Processing of DNA double-strand break ends / double-stranded DNA binding / DNA-binding transcription factor binding / DNA recombination / cell population proliferation / Regulation of TP53 Activity through Phosphorylation / damaged DNA binding / chromosome, telomeric region / Hydrolases; Acting on ester bonds / regulation of cell cycle / cadherin binding / DNA repair / DNA damage response / nucleolus / negative regulation of apoptotic process / Golgi apparatus / nucleoplasm / identical protein binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Homo sapiens (human) | |||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.13 Å | |||||||||||||||
Authors | Bartho, J.D. / Rotheneder, M. / Stakyte, K. / Lammens, K. / Hopfner, K.P. | |||||||||||||||
Funding support | Germany, 4items
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Citation | Journal: Mol Cell / Year: 2023 Title: Cryo-EM structure of the Mre11-Rad50-Nbs1 complex reveals the molecular mechanism of scaffolding functions. Authors: Matthias Rotheneder / Kristina Stakyte / Erik van de Logt / Joseph D Bartho / Katja Lammens / Yilan Fan / Aaron Alt / Brigitte Kessler / Christophe Jung / Wynand P Roos / Barbara ...Authors: Matthias Rotheneder / Kristina Stakyte / Erik van de Logt / Joseph D Bartho / Katja Lammens / Yilan Fan / Aaron Alt / Brigitte Kessler / Christophe Jung / Wynand P Roos / Barbara Steigenberger / Karl-Peter Hopfner / Abstract: The DNA double-strand break repair complex Mre11-Rad50-Nbs1 (MRN) detects and nucleolytically processes DNA ends, activates the ATM kinase, and tethers DNA at break sites. How MRN can act both as ...The DNA double-strand break repair complex Mre11-Rad50-Nbs1 (MRN) detects and nucleolytically processes DNA ends, activates the ATM kinase, and tethers DNA at break sites. How MRN can act both as nuclease and scaffold protein is not well understood. The cryo-EM structure of MRN from Chaetomium thermophilum reveals a 2:2:1 complex with a single Nbs1 wrapping around the autoinhibited Mre11 nuclease dimer. MRN has two DNA-binding modes, one ATP-dependent mode for loading onto DNA ends and one ATP-independent mode through Mre11's C terminus, suggesting how it may interact with DSBs and intact DNA. MRNs two 60-nm-long coiled-coil domains form a linear rod structure, the apex of which is assembled by the two joined zinc-hook motifs. Apices from two MRN complexes can further dimerize, forming 120-nm spanning MRN-MRN structures. Our results illustrate the architecture of MRN and suggest how it mechanistically integrates catalytic and tethering functions. | |||||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8bah.cif.gz | 316.3 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8bah.ent.gz | 244 KB | Display | PDB format |
PDBx/mmJSON format | 8bah.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ba/8bah ftp://data.pdbj.org/pub/pdb/validation_reports/ba/8bah | HTTPS FTP |
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-Related structure data
Related structure data | 15948MC 7zqyC 7zr1C M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 84008.633 Da / Num. of mol.: 2 / Mutation: H129N Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: MRE11, HNGS1, MRE11A / Plasmid: pACEMam1_pMDC / Cell line (production host): HEK293T / Production host: Homo sapiens (human) References: UniProt: P49959, Hydrolases; Acting on ester bonds #2: Protein | | Mass: 85073.023 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NBN, NBS, NBS1, P95 / Plasmid: pACEMam1_pMDC / Cell line (production host): HEK293T / Production host: Homo sapiens (human) / References: UniProt: O60934 #3: Chemical | ChemComp-MN / Has ligand of interest | Y | |
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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 |
-Sample preparation
Component | Name: Human Mre11-Nbs1 complex / Type: COMPLEX Details: Human Mre11-dimer with Nbs1 C-terminal chain bound. Entity ID: #1-#2 / Source: RECOMBINANT | |||||||||||||||||||||||||||||||||||||||||||||
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Molecular weight | Experimental value: NO | |||||||||||||||||||||||||||||||||||||||||||||
Source (natural) | Organism: Homo sapiens (human) | |||||||||||||||||||||||||||||||||||||||||||||
Source (recombinant) | Organism: Homo sapiens (human) / Cell: HEK293T / Plasmid: pACEMam1_pMDC | |||||||||||||||||||||||||||||||||||||||||||||
Buffer solution | pH: 7 Details: 20 mM HEPES (pH 7.0), 140 mM NaCl, 5 mM MgCl2, 1 mM MnCl2, 0.020 mM ZnCl2, 0.2 mM TCEP, 2 mM ATPgS, plus 0.05 percent beta-OG | |||||||||||||||||||||||||||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 0.29 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | |||||||||||||||||||||||||||||||||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R2/1 | |||||||||||||||||||||||||||||||||||||||||||||
Vitrification | Instrument: LEICA EM GP / Cryogen name: ETHANE / Humidity: 95 % / Chamber temperature: 288 K |
-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: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELDBright-field microscopy / Nominal magnification: 13000 X / Nominal defocus max: 2800 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 10 sec. / Electron dose: 43 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 3 / Num. of real images: 11325 |
EM imaging optics | Energyfilter name: GIF Bioquantum / Energyfilter slit width: 20 eV |
Image scans | Movie frames/image: 40 |
-Processing
Software | Name: PHENIX / Version: 1.20.1_4487: / Classification: refinement | ||||||||||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 4.13 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 282838 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
Refine LS restraints |
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