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Open data
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Basic information
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| Title | sCMGE assembled on ARS1 DNA with Sld2 and RPA | |||||||||||||||
Map data | Final refinement map, unmodified. | |||||||||||||||
Sample |
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Keywords | Macromolecular complex / DNA / ATPase / Helicase MCM2-7 / REPLICATION | |||||||||||||||
| Function / homology | Function and homology informationgene conversion / DNA-templated DNA replication maintenance of fidelity / Unwinding of DNA / DNA replication initiation / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / GINS complex ...gene conversion / DNA-templated DNA replication maintenance of fidelity / Unwinding of DNA / DNA replication initiation / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / GINS complex / MCM complex binding / mitotic DNA replication preinitiation complex assembly / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nucleotide-excision repair, DNA gap filling / SUMO binding / Activation of the pre-replicative complex / mitotic DNA replication / nuclear pre-replicative complex / CMG complex / DNA replication proofreading / : / DNA replication preinitiation complex / single-stranded DNA 3'-5' DNA exonuclease activity / Activation of ATR in response to replication stress / mitotic DNA replication checkpoint signaling / double-strand break repair via break-induced replication / MCM complex / mitotic DNA replication initiation / mitotic intra-S DNA damage checkpoint signaling / silent mating-type cassette heterochromatin formation / single-stranded DNA helicase activity / regulation of DNA-templated DNA replication initiation / mitotic sister chromatid cohesion / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / DNA strand elongation involved in DNA replication / leading strand elongation / nuclear replication fork / DNA replication origin binding / Dual incision in TC-NER / error-prone translesion synthesis / DNA replication initiation / subtelomeric heterochromatin formation / base-excision repair, gap-filling / replication fork / DNA helicase activity / helicase activity / transcription elongation by RNA polymerase II / base-excision repair / DNA-templated DNA replication / double-strand break repair via nonhomologous end joining / peroxisome / double-strand break repair / mitotic cell cycle / heterochromatin formation / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / double-stranded DNA binding / DNA-directed DNA polymerase / DNA helicase / DNA-directed DNA polymerase activity / DNA replication / chromosome, telomeric region / nucleotide binding / mRNA binding / chromatin binding / DNA damage response / ATP hydrolysis activity / DNA binding / zinc ion binding / nucleoplasm / ATP binding / nucleus / cytoplasm Similarity search - Function | |||||||||||||||
| Biological species | ![]() | |||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.7 Å | |||||||||||||||
Authors | Palm G / Costa A | |||||||||||||||
| Funding support | United Kingdom, European Union, Germany, 4 items
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Citation | Journal: Nature / Year: 2026Title: Structure of the pre-initiation complex explains CMGE biogenesis. Authors: Thomas Pühringer / Berta Canal / Giacomo Palm / Agata Butryn / Emma C Couves / Oliver Willhoft / Jacob S Lewis / John F X Diffley / Alessandro Costa / ![]() Abstract: When cells enter S phase, bidirectional DNA replication is initiated through the kinase-regulated recruitment of three activators (Cdc45, GINS and Pol ε) to a duplex-DNA-loaded double hexamer of ...When cells enter S phase, bidirectional DNA replication is initiated through the kinase-regulated recruitment of three activators (Cdc45, GINS and Pol ε) to a duplex-DNA-loaded double hexamer of minichromosome maintenance (MCM) ATPases. Together, these proteins form two CMGE helicases that establish divergent replication forks as they become separated. Here, to gain an understanding of CMGE biogenesis, we reconstituted the pre-initiation complex with purified yeast proteins. The cryo-electron-microscopy structure shows a set of firing factors caught in the act of assembling two symmetrical CMGEs. We show how stepwise complex formation reshapes MCM in preparation for DNA opening, and we explain how ATP promotes firing-factor ejection and CMGE maturation. We find that although Sld2 facilitates the recruitment of GINS to MCM, as expected, it also aids the efficient separation of the CMGE dimer, and is essential for the ejection of the lagging strand from MCM. These findings have direct implications for our understanding of the metazoan Sld2 orthologue, RECQL4, and point to a replication-fork establishment mechanism that is conserved across eukaryotes. | |||||||||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_56898.map.gz | 154.9 MB | EMDB map data format | |
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| Header (meta data) | emd-56898-v30.xml emd-56898.xml | 47 KB 47 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_56898_fsc.xml | 14.2 KB | Display | FSC data file |
| Images | emd_56898.png | 62.1 KB | ||
| Filedesc metadata | emd-56898.cif.gz | 13.9 KB | ||
| Others | emd_56898_additional_1.map.gz emd_56898_half_map_1.map.gz emd_56898_half_map_2.map.gz | 196.3 MB 285.6 MB 285.6 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-56898 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-56898 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 28vyMC ![]() 9rhiC ![]() 9rhjC ![]() 9rhlC ![]() 9rhmC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_56898.map.gz / Format: CCP4 / Size: 307.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Final refinement map, unmodified. | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.95 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: Final refinement map, denisty modified with EMReady.
| File | emd_56898_additional_1.map | ||||||||||||
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| Annotation | Final refinement map, denisty modified with EMReady. | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: Final refinement half map
| File | emd_56898_half_map_1.map | ||||||||||||
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| Annotation | Final refinement half map | ||||||||||||
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| Density Histograms |
-Half map: Final refinement half map
| File | emd_56898_half_map_2.map | ||||||||||||
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| Annotation | Final refinement half map | ||||||||||||
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| Density Histograms |
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Sample components
+Entire : Single CMGE complex with RPA on replication origin
+Supramolecule #1: Single CMGE complex with RPA on replication origin
+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 #8: DNA replication complex GINS protein PSF3
+Macromolecule #9: DNA replication complex GINS protein SLD5
+Macromolecule #10: Cell division control protein 45
+Macromolecule #11: DNA polymerase epsilon subunit B
+Macromolecule #12: DNA polymerase epsilon catalytic subunit A
+Macromolecule #13: DNA replication complex GINS protein PSF1
+Macromolecule #14: DNA replication complex GINS protein PSF2
+Macromolecule #7: DNA (5'-D(P*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP*TP*CP*GP*AP...
+Macromolecule #15: MAGNESIUM ION
+Macromolecule #16: ZINC ION
+Macromolecule #17: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #18: ADENOSINE-5'-TRIPHOSPHATE
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.6 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 38.6 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.9 µm / Nominal defocus min: 2.0 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi




Keywords
Authors
United Kingdom, European Union,
Germany, 4 items
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Processing
FIELD EMISSION GUN

