+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10290 | |||||||||
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Title | Structure of Fanconi anaemia core complex (consensus map) | |||||||||
Map data | This is the post processed consensus map. To see the fit of model into map, please use the focussed maps for top, middle, bottom and superimposed consensus maps with and without T=5 on middle map. | |||||||||
Sample |
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Keywords | Fanconi Anaemia core complex / E3 ligase / DNA repair / LIGASE | |||||||||
Function / homology | Function and homology information PKR-mediated signaling / Fanconi Anemia Pathway in DNA repair / Fanconi Anemia Pathway / Fanconi anaemia nuclear complex / protein monoubiquitination / interstrand cross-link repair / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / DNA repair / DNA damage response ...PKR-mediated signaling / Fanconi Anemia Pathway in DNA repair / Fanconi Anemia Pathway / Fanconi anaemia nuclear complex / protein monoubiquitination / interstrand cross-link repair / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / DNA repair / DNA damage response / nucleoplasm / nucleus Similarity search - Function | |||||||||
Biological species | Gallus gallus (chicken) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.2 Å | |||||||||
Authors | Shakeel S / Rajendra E | |||||||||
Funding support | United Kingdom, 1 items
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Citation | Journal: Nature / Year: 2019 Title: Structure of the Fanconi anaemia monoubiquitin ligase complex. Authors: Shabih Shakeel / Eeson Rajendra / Pablo Alcón / Francis O'Reilly / Dror S Chorev / Sarah Maslen / Gianluca Degliesposti / Christopher J Russo / Shaoda He / Chris H Hill / J Mark Skehel / ...Authors: Shabih Shakeel / Eeson Rajendra / Pablo Alcón / Francis O'Reilly / Dror S Chorev / Sarah Maslen / Gianluca Degliesposti / Christopher J Russo / Shaoda He / Chris H Hill / J Mark Skehel / Sjors H W Scheres / Ketan J Patel / Juri Rappsilber / Carol V Robinson / Lori A Passmore / Abstract: The Fanconi anaemia (FA) pathway repairs DNA damage caused by endogenous and chemotherapy-induced DNA crosslinks, and responds to replication stress. Genetic inactivation of this pathway by mutation ...The Fanconi anaemia (FA) pathway repairs DNA damage caused by endogenous and chemotherapy-induced DNA crosslinks, and responds to replication stress. Genetic inactivation of this pathway by mutation of genes encoding FA complementation group (FANC) proteins impairs development, prevents blood production and promotes cancer. The key molecular step in the FA pathway is the monoubiquitination of a pseudosymmetric heterodimer of FANCD2-FANCI by the FA core complex-a megadalton multiprotein E3 ubiquitin ligase. Monoubiquitinated FANCD2 then recruits additional protein factors to remove the DNA crosslink or to stabilize the stalled replication fork. A molecular structure of the FA core complex would explain how it acts to maintain genome stability. Here we reconstituted an active, recombinant FA core complex, and used cryo-electron microscopy and mass spectrometry to determine its structure. The FA core complex comprises two central dimers of the FANCB and FA-associated protein of 100 kDa (FAAP100) subunits, flanked by two copies of the RING finger subunit, FANCL. These two heterotrimers act as a scaffold to assemble the remaining five subunits, resulting in an extended asymmetric structure. Destabilization of the scaffold would disrupt the entire complex, resulting in a non-functional FA pathway. Thus, the structure provides a mechanistic basis for the low numbers of patients with mutations in FANCB, FANCL and FAAP100. Despite a lack of sequence homology, FANCB and FAAP100 adopt similar structures. The two FANCL subunits are in different conformations at opposite ends of the complex, suggesting that each FANCL has a distinct role. This structural and functional asymmetry of dimeric RING finger domains may be a general feature of E3 ligases. The cryo-electron microscopy structure of the FA core complex provides a foundation for a detailed understanding of its E3 ubiquitin ligase activity and DNA interstrand crosslink repair. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_10290.map.gz | 38.1 MB | EMDB map data format | |
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Header (meta data) | emd-10290-v30.xml emd-10290.xml | 60.8 KB 60.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_10290_fsc.xml | 20.8 KB | Display | FSC data file |
Images | emd_10290.png | 66.8 KB | ||
Masks | emd_10290_msk_1.map | 767.6 MB | Mask map | |
Filedesc metadata | emd-10290.cif.gz | 7.2 KB | ||
Others | emd_10290_additional_1.map.gz emd_10290_additional_2.map.gz emd_10290_additional_3.map.gz emd_10290_half_map_1.map.gz emd_10290_half_map_2.map.gz | 620.3 MB 620.3 MB 620.3 MB 622.1 MB 621.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10290 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10290 | HTTPS FTP |
-Validation report
Summary document | emd_10290_validation.pdf.gz | 635.1 KB | Display | EMDB validaton report |
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Full document | emd_10290_full_validation.pdf.gz | 634.7 KB | Display | |
Data in XML | emd_10290_validation.xml.gz | 30 KB | Display | |
Data in CIF | emd_10290_validation.cif.gz | 38.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10290 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10290 | HTTPS FTP |
-Related structure data
Related structure data | 6sriMC 6srsC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | |
EM raw data | EMPIAR-10608 (Title: Unaligned movies for FA core complex and symmetric complex Data size: 13.2 TB Data #1: Unaligned movies for FA core complex and symmetric complex [micrographs - multiframe] Data #2: Unaligned movies for FA core complex and symmetric complex [micrographs - multiframe]) |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_10290.map.gz / Format: CCP4 / Size: 767.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | This is the post processed consensus map. To see the fit of model into map, please use the focussed maps for top, middle, bottom and superimposed consensus maps with and without T=5 on middle map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.04 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_10290_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: This is the map from auto refinement in Relion
File | emd_10290_additional_1.map | ||||||||||||
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Annotation | This is the map from auto refinement in Relion | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Consensus map refined with T=5 value in Relion,...
File | emd_10290_additional_2.map | ||||||||||||
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Annotation | Consensus map refined with T=5 value in Relion, post processed and superimposed on middle map using Proshade from ccpem. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: This is the map from auto refinement in Relion.
File | emd_10290_additional_3.map | ||||||||||||
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Annotation | This is the map from auto refinement in Relion. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: This is half1 map from auto refinement in Relion.
File | emd_10290_half_map_1.map | ||||||||||||
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Annotation | This is half1 map from auto refinement in Relion. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: This is half2 map from auto refinement in Relion.
File | emd_10290_half_map_2.map | ||||||||||||
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Annotation | This is half2 map from auto refinement in Relion. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Fanconi anaemia core complex
+Supramolecule #1: Fanconi anaemia core complex
+Macromolecule #1: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #2: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #3: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #4: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #5: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #6: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #7: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #8: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #9: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #10: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #11: Unassigned secondary structure elements (central region, proposed...
+Macromolecule #12: Unassigned secondary structure elements (proposed FANCB)
+Macromolecule #13: Unassigned secondary structure elements (proposed FAAP100)
+Macromolecule #14: Unassigned secondary structure elements (proposed FANCB)
+Macromolecule #15: Unassigned secondary structure elements (proposed FAAP100)
+Macromolecule #16: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #17: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #18: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #19: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #20: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #21: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #22: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #23: Unassigned secondary structure elements (base region, proposed FA...
+Macromolecule #24: Fanconi anaemia protein FANCL
+Macromolecule #25: base region, proposed FANCF
+Macromolecule #26: Unassigned secondary structure elements (top region, proposed FANCG)
+Macromolecule #27: Unassigned secondary structure elements (top region, proposed FANCG)
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.7 mg/mL |
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Buffer | pH: 8 / Details: 50 mM HEPES pH 8.0, ~500 mM NaCl, 1 mM TCEP |
Grid | Model: Quantifoil, UltrAuFoil, R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - topology: HOLEY / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 30 sec. / Details: Argon: 9 parts Oxygen: 1 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV Details: Blot time: 3 to 4.5 s Wait time: 0 s Drain time: 0 s Force: -10 N. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Number real images: 4145 / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: -4.0 µm / Nominal defocus min: -1.8 µm / Nominal magnification: 75000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Protocol: OTHER |
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Output model | PDB-6sri: |