+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-17449 | |||||||||||||||
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Title | S. cerevisiae nexus-sCMGE after DNA replication initiation | |||||||||||||||
Map data | EM map of nexus-sCMGE | |||||||||||||||
Sample |
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Keywords | Saccharomyces cerevisiae / helicase / CMGE / initiation of DNA replication / DNA / DNA unwinding / REPLICATION | |||||||||||||||
Function / homology | Function and homology information DNA-templated DNA replication maintenance of fidelity / gene conversion / Unwinding of DNA / DNA replication initiation / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / MCM complex binding ...DNA-templated DNA replication maintenance of fidelity / gene conversion / Unwinding of DNA / DNA replication initiation / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / MCM complex binding / GINS complex / nuclear DNA replication / mitotic DNA replication preinitiation complex assembly / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / SUMO binding / nucleotide-excision repair, DNA gap filling / mitotic DNA replication / Activation of the pre-replicative complex / DNA replication proofreading / CMG complex / nuclear pre-replicative complex / single-stranded DNA 3'-5' DNA exonuclease activity / Activation of ATR in response to replication stress / DNA replication preinitiation complex / MCM complex / replication fork protection complex / Termination of translesion DNA synthesis / 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 / mitotic intra-S DNA damage checkpoint signaling / DNA strand elongation involved in DNA replication / silent mating-type cassette heterochromatin formation / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / mitotic sister chromatid cohesion / leading strand elongation / DNA unwinding involved in DNA replication / nuclear replication fork / DNA replication origin binding / Dual incision in TC-NER / DNA replication initiation / subtelomeric heterochromatin formation / error-prone translesion synthesis / base-excision repair, gap-filling / DNA helicase activity / replication fork / helicase activity / transcription elongation by RNA polymerase II / base-excision repair / heterochromatin formation / DNA-templated DNA replication / double-strand break repair via nonhomologous end joining / double-strand break repair / mitotic cell cycle / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / double-stranded DNA binding / DNA helicase / DNA-directed DNA polymerase / chromosome, telomeric region / DNA-directed DNA polymerase activity / mRNA binding / nucleotide binding / DNA damage response / chromatin binding / ATP hydrolysis activity / DNA binding / zinc ion binding / nucleoplasm / ATP binding / nucleus / metal ion binding / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) / synthetic construct (others) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||||||||
Authors | Henrikus SS / Willhoft O | |||||||||||||||
Funding support | United Kingdom, European Union, France, 4 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2024 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 | emd_17449.map.gz | 18.3 MB | EMDB map data format | |
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Header (meta data) | emd-17449-v30.xml emd-17449.xml | 44.3 KB 44.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_17449_fsc.xml | 14.9 KB | Display | FSC data file |
Images | emd_17449.png | 172.3 KB | ||
Filedesc metadata | emd-17449.cif.gz | 13.5 KB | ||
Others | emd_17449_half_map_1.map.gz emd_17449_half_map_2.map.gz | 225.3 MB 225.2 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-17449 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-17449 | HTTPS FTP |
-Validation report
Summary document | emd_17449_validation.pdf.gz | 923.9 KB | Display | EMDB validaton report |
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Full document | emd_17449_full_validation.pdf.gz | 923.4 KB | Display | |
Data in XML | emd_17449_validation.xml.gz | 22.5 KB | Display | |
Data in CIF | emd_17449_validation.cif.gz | 29.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-17449 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-17449 | HTTPS FTP |
-Related structure data
Related structure data | 8p5eMC 8p62C 8p63C M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_17449.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | EM map of nexus-sCMGE | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.08 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: EM half map 1 of nexus-sCMGE
File | emd_17449_half_map_1.map | ||||||||||||
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Annotation | EM half map 1 of nexus-sCMGE | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: EM half map 2 of nexus-sCMGE
File | emd_17449_half_map_2.map | ||||||||||||
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Annotation | EM half map 2 of nexus-sCMGE | ||||||||||||
Projections & Slices |
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Density Histograms |
-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: Minichromosome maintenance protein 5
+Macromolecule #5: DNA replication licensing factor MCM6
+Macromolecule #6: DNA replication licensing factor MCM7
+Macromolecule #9: DNA replication complex GINS protein PSF3
+Macromolecule #10: DNA replication complex GINS protein SLD5
+Macromolecule #11: Cell division control protein 45
+Macromolecule #12: DNA polymerase epsilon subunit B
+Macromolecule #13: DNA polymerase epsilon catalytic subunit A
+Macromolecule #14: DNA replication complex GINS protein PSF1
+Macromolecule #15: DNA replication complex GINS protein PSF2
+Macromolecule #7: DNA (19-MER)
+Macromolecule #8: DNA (7-MER)
+Macromolecule #16: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #17: ZINC ION
+Macromolecule #18: MAGNESIUM ION
+Macromolecule #19: ADENOSINE-5'-DIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.6 |
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Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
-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: FIELD EMISSION GUN |
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 |