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Yorodumi- EMDB-8540: Structural basis of MCM2-7 replicative helicase loading by ORC-Cd... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-8540 | |||||||||
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Title | Structural basis of MCM2-7 replicative helicase loading by ORC-Cdc6 and Cdt1 | |||||||||
Map data | Tetrameric HIV-1 Strand Transfer Complex Intasome | |||||||||
Sample |
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Function / homology | Function and homology information MCM complex loading / CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / Assembly of the ORC complex at the origin of replication / nuclear DNA replication ...MCM complex loading / CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / Assembly of the ORC complex at the origin of replication / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear origin of replication recognition complex / mitotic DNA replication / Activation of the pre-replicative complex / nucleosome organization / CMG complex / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / MCM complex / cyclin-dependent protein serine/threonine kinase inhibitor activity / replication fork protection complex / mitotic DNA replication checkpoint signaling / double-strand break repair via break-induced replication / mitotic DNA replication initiation / single-stranded DNA helicase activity / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / silent mating-type cassette heterochromatin formation / CDK-mediated phosphorylation and removal of Cdc6 / DNA unwinding involved in DNA replication / Orc1 removal from chromatin / nuclear replication fork / regulation of DNA replication / DNA replication origin binding / nucleosome binding / DNA replication initiation / subtelomeric heterochromatin formation / DNA helicase activity / helicase activity / transcription elongation by RNA polymerase II / heterochromatin formation / G1/S transition of mitotic cell cycle / single-stranded DNA binding / chromosome / DNA helicase / chromosome, telomeric region / cell division / GTPase activity / DNA damage response / chromatin binding / GTP binding / ATP hydrolysis activity / nucleoplasm / ATP binding / nucleus / metal ion binding / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) / Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) / synthetic construct (others) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||
Authors | Yuan Z / Riera A / Bai L / Sun J / Spanos C / Chen ZA / Barbon M / Rappsilber J / Stillman B / Speck C / Li H | |||||||||
Citation | Journal: Nat Struct Mol Biol / Year: 2017 Title: Structural basis of Mcm2-7 replicative helicase loading by ORC-Cdc6 and Cdt1. Authors: Zuanning Yuan / Alberto Riera / Lin Bai / Jingchuan Sun / Saikat Nandi / Christos Spanos / Zhuo Angel Chen / Marta Barbon / Juri Rappsilber / Bruce Stillman / Christian Speck / Huilin Li / Abstract: To initiate DNA replication, the origin recognition complex (ORC) and Cdc6 load an Mcm2-7 double hexamer onto DNA. Without ATP hydrolysis, ORC-Cdc6 recruits one Cdt1-bound Mcm2-7 hexamer, thus ...To initiate DNA replication, the origin recognition complex (ORC) and Cdc6 load an Mcm2-7 double hexamer onto DNA. Without ATP hydrolysis, ORC-Cdc6 recruits one Cdt1-bound Mcm2-7 hexamer, thus forming an ORC-Cdc6-Cdt1-Mcm2-7 (OCCM) helicase-loading intermediate. Here we report a 3.9-Å structure of Saccharomyces cerevisiae OCCM on DNA. Flexible Mcm2-7 winged-helix domains (WHDs) engage ORC-Cdc6. A three-domain Cdt1 configuration embraces Mcm2, Mcm4, and Mcm6, thus comprising nearly half of the hexamer. The Cdt1 C-terminal domain extends to the Mcm6 WHD, which binds the Orc4 WHD. DNA passes through the ORC-Cdc6 and Mcm2-7 rings. Origin DNA interaction is mediated by an α-helix within Orc4 and positively charged loops within Orc2 and Cdc6. The Mcm2-7 C-tier AAA+ ring is topologically closed by an Mcm5 loop that embraces Mcm2, but the N-tier-ring Mcm2-Mcm5 interface remains open. This structure suggests a loading mechanism of the first Cdt1-bound Mcm2-7 hexamer by ORC-Cdc6. | |||||||||
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_8540.map.gz | 58.9 MB | EMDB map data format | |
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Header (meta data) | emd-8540-v30.xml emd-8540.xml | 35.8 KB 35.8 KB | Display Display | EMDB header |
Images | emd_8540.png | 152.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-8540 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8540 | HTTPS FTP |
-Validation report
Summary document | emd_8540_validation.pdf.gz | 473 KB | Display | EMDB validaton report |
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Full document | emd_8540_full_validation.pdf.gz | 472.6 KB | Display | |
Data in XML | emd_8540_validation.xml.gz | 5.8 KB | Display | |
Data in CIF | emd_8540_validation.cif.gz | 6.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8540 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8540 | HTTPS FTP |
-Related structure data
Related structure data | 5v8fMC 5udb M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_8540.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Tetrameric HIV-1 Strand Transfer Complex Intasome | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.01 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 : ORC-Cdc6-Cdt1-MCM2-7
+Supramolecule #1: ORC-Cdc6-Cdt1-MCM2-7
+Macromolecule #1: DNA replication licensing factor MCM2
+Macromolecule #2: DNA replication licensing factor MCM3
+Macromolecule #3: DNA replication licensing factor MCM4
+Macromolecule #4: Minichromosome maintenance protein 5
+Macromolecule #5: DNA replication licensing factor MCM6
+Macromolecule #6: DNA replication licensing factor MCM7
+Macromolecule #7: Cell division cycle protein CDT1
+Macromolecule #8: Cell division control protein 6
+Macromolecule #9: Origin recognition complex subunit 1
+Macromolecule #10: Origin recognition complex subunit 2
+Macromolecule #11: Origin recognition complex subunit 3
+Macromolecule #12: Origin recognition complex subunit 4
+Macromolecule #13: Origin recognition complex subunit 5
+Macromolecule #14: Origin recognition complex subunit 6
+Macromolecule #15: DNA (39-MER)
+Macromolecule #16: DNA (39-MER)
+Macromolecule #17: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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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: 50.0 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: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 304288 |
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Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |
-Atomic model buiding 1
Refinement | Protocol: RIGID BODY FIT |
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Output model | PDB-5v8f: |