+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11021 | ||||||||||||
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Title | CTC-DNA2, body 2 (contains MMS19 and CIAO2B) | ||||||||||||
Map data | |||||||||||||
Sample |
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Biological species | Homo sapiens (human) / Mus musculus (house mouse) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 12.2 Å | ||||||||||||
Authors | Kassube SA / Thomae N | ||||||||||||
Funding support | Switzerland, 3 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2020 Title: Structural insights into Fe-S protein biogenesis by the CIA targeting complex. Authors: Susanne A Kassube / Nicolas H Thomä / Abstract: The cytosolic iron-sulfur (Fe-S) assembly (CIA) pathway is required for the insertion of Fe-S clusters into cytosolic and nuclear client proteins, including many DNA replication and repair factors. ...The cytosolic iron-sulfur (Fe-S) assembly (CIA) pathway is required for the insertion of Fe-S clusters into cytosolic and nuclear client proteins, including many DNA replication and repair factors. The molecular mechanisms of client protein recognition and Fe-S cluster transfer remain unknown. Here, we report crystal structures of the CIA targeting complex (CTC), revealing that its CIAO2B subunit is centrally located and bridges CIAO1 and the client adaptor protein MMS19. Cryo-EM reconstructions of human CTC bound either to the DNA replication factor primase or to the DNA helicase DNA2, combined with biochemical, biophysical and yeast complementation assays, reveal an evolutionarily conserved, bipartite client recognition mode facilitated by CIAO1 and the structural flexibility of the MMS19 subunit. Unexpectedly, the primase Fe-S cluster is located ~70 Å away from the CTC reactive cysteine, implicating conformational dynamics of the CTC or additional maturation factors in the mechanism of Fe-S cluster transfer. | ||||||||||||
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_11021.map.gz | 3.6 MB | EMDB map data format | |
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Header (meta data) | emd-11021-v30.xml emd-11021.xml | 9.5 KB 9.5 KB | Display Display | EMDB header |
Images | emd_11021.png | 27.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-11021 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11021 | HTTPS FTP |
-Validation report
Summary document | emd_11021_validation.pdf.gz | 187.2 KB | Display | EMDB validaton report |
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Full document | emd_11021_full_validation.pdf.gz | 186.2 KB | Display | |
Data in XML | emd_11021_validation.xml.gz | 6.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11021 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11021 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_11021.map.gz / Format: CCP4 / Size: 91.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Voxel size | X=Y=Z: 1.1 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : MMS19-CIAO2B complex
Entire | Name: MMS19-CIAO2B complex |
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Components |
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-Supramolecule #1: MMS19-CIAO2B complex
Supramolecule | Name: MMS19-CIAO2B complex / type: complex / ID: 1 / Parent: 0 |
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Molecular weight | Theoretical: 119.447 KDa |
-Supramolecule #2: MMS19-CIAO1-CIAO2B CIA targeting complex
Supramolecule | Name: MMS19-CIAO1-CIAO2B CIA targeting complex / type: complex / ID: 2 / Parent: 1 |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
-Supramolecule #3: DNA2
Supramolecule | Name: DNA2 / type: complex / ID: 3 / Parent: 1 |
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Source (natural) | Organism: Mus musculus (house mouse) |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.4 mg/mL |
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Buffer | pH: 7.4 |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 39.9 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 12.2 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 69894 |
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Initial angle assignment | Type: OTHER |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |