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Open data
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Basic information
Entry | ![]() | |||||||||||||||
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Title | S. cerevisiae sCMGE with N-ter Mcm10 density | |||||||||||||||
![]() | EM map of sCMG10 | |||||||||||||||
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![]() | Saccharomyces cerevisiae / helicase / CMGE / initiation of DNA replication / DNA / DNA unwinding / REPLICATION / Mcm10 | |||||||||||||||
Biological species | ![]() ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.5 Å | |||||||||||||||
![]() | Henrikus SS / Willhoft O | |||||||||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Unwinding of a eukaryotic origin of replication visualized by cryo-EM. Authors: Sarah S Henrikus / Marta H Gross / Oliver Willhoft / Thomas Pühringer / Jacob S Lewis / Allison W McClure / Julia F Greiwe / Giacomo Palm / Andrea Nans / John F X Diffley / Alessandro Costa / ![]() ![]() ![]() Abstract: To prevent detrimental chromosome re-replication, DNA loading of a double hexamer of the minichromosome maintenance (MCM) replicative helicase is temporally separated from DNA unwinding. Upon S-phase ...To prevent detrimental chromosome re-replication, DNA loading of a double hexamer of the minichromosome maintenance (MCM) replicative helicase is temporally separated from DNA unwinding. Upon S-phase transition in yeast, DNA unwinding is achieved in two steps: limited opening of the double helix and topological separation of the two DNA strands. First, Cdc45, GINS and Polε engage MCM to assemble a double CMGE with two partially separated hexamers that nucleate DNA melting. In the second step, triggered by Mcm10, two CMGEs separate completely, eject the lagging-strand template and cross paths. To understand Mcm10 during helicase activation, we used biochemical reconstitution with cryogenic electron microscopy. We found that Mcm10 splits the double CMGE by engaging the N-terminal homo-dimerization face of MCM. To eject the lagging strand, DNA unwinding is started from the N-terminal side of MCM while the hexamer channel becomes too narrow to harbor duplex DNA. | |||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 2.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 33.8 KB 33.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 6.4 KB | Display | ![]() |
Images | ![]() | 82.4 KB | ||
Filedesc metadata | ![]() | 9.2 KB | ||
Others | ![]() ![]() | 17 MB 17 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 682 KB | Display | ![]() |
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Full document | ![]() | 681.5 KB | Display | |
Data in XML | ![]() | 11.9 KB | Display | |
Data in CIF | ![]() | 16.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||
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Annotation | EM map of sCMG10 | ||||||||||||||||||||
Voxel size | X=Y=Z: 2.16 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: EM half map 2 of sCMG10
File | emd_17460_half_map_1.map | ||||||||||||
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Annotation | EM half map 2 of sCMG10 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: EM half map 1 of sCMG10
File | emd_17460_half_map_2.map | ||||||||||||
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Annotation | EM half map 1 of sCMG10 | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
+Entire : Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsi...
+Supramolecule #1: Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsi...
+Supramolecule #2: DNA replication complex GINS containing Psf1, Psf2, Psf3 and Sld5
+Supramolecule #3: DNA replication licensing factors MCM2-7
+Supramolecule #4: DNA polymerase epsilon contains catalytic subunit A and non-catal...
+Supramolecule #5: DNA
+Supramolecule #6: Cdc45
+Macromolecule #1: DNA replication licensing factor MCM2
+Macromolecule #2: DNA replication licensing factor MCM3
+Macromolecule #3: DNA replication licensing factor MCM4
+Macromolecule #4: DNA replication licensing factor MCM5
+Macromolecule #5: DNA replication licensing factor MCM6
+Macromolecule #6: Cell division control protein 45
+Macromolecule #7: DNA replication complex GINS protein PSF1
+Macromolecule #8: DNA replication complex GINS protein PSF2
+Macromolecule #9: DNA replication complex GINS protein PSF3
+Macromolecule #10: DNA replication complex GINS protein SLD5
+Macromolecule #11: Minichromosome maintenance protein 10
+Macromolecule #14: DNA replication licensing factor MCM7
+Macromolecule #12: DNA strand 1
+Macromolecule #13: DNA strand 2
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.6 |
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Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
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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: 1.67 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.3000000000000003 µm / Nominal defocus min: 1.8 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |