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Open data
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Basic information
| Entry | ![]() | |||||||||||||||||||||
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| Title | Phospho-DH bound by Sld3-MBD on ARS1 DNA | |||||||||||||||||||||
Map data | Local Refinement of DH-MBD cryoSPARC Unsharpened map | |||||||||||||||||||||
Sample |
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Keywords | Macromolecular Complex / DNA / ATPase / Helicase / MCM2-7 / REPLICATION | |||||||||||||||||||||
| Function / homology | Function and homology informationregulation of mitotic DNA replication initiation / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / mitotic DNA replication ...regulation of mitotic DNA replication initiation / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / mitotic DNA replication / nuclear pre-replicative complex / CMG complex / DNA replication preinitiation complex / 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 / silent mating-type cassette heterochromatin formation / single-stranded DNA helicase activity / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / nuclear replication fork / DNA replication origin binding / chromosome, centromeric region / DNA replication initiation / subtelomeric heterochromatin formation / DNA helicase activity / helicase activity / transcription elongation by RNA polymerase II / peroxisome / heterochromatin formation / single-stranded DNA binding / DNA helicase / DNA replication / chromosome, telomeric region / chromatin binding / DNA damage response / chromatin / ATP hydrolysis activity / zinc ion binding / nucleoplasm / ATP binding / nucleus / cytoplasm Similarity search - Function | |||||||||||||||||||||
| Biological species | ![]() | |||||||||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||||||||||||||
Authors | Puehringer T / Couves EC / Costa A | |||||||||||||||||||||
| Funding support | United Kingdom, European Union, Germany, 6 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
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_53972.map.gz | 172.1 MB | EMDB map data format | |
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| Header (meta data) | emd-53972-v30.xml emd-53972.xml | 39.5 KB 39.5 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_53972_fsc.xml | 14.7 KB | Display | FSC data file |
| Images | emd_53972.png | 67.3 KB | ||
| Filedesc metadata | emd-53972.cif.gz | 11.4 KB | ||
| Others | emd_53972_additional_1.map.gz emd_53972_half_map_1.map.gz emd_53972_half_map_2.map.gz | 256 MB 317.7 MB 317.7 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-53972 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-53972 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9rhlMC ![]() 28vyC ![]() 9rhiC ![]() 9rhjC ![]() 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_53972.map.gz / Format: CCP4 / Size: 343 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Local Refinement of DH-MBD cryoSPARC Unsharpened map | ||||||||||||||||||||||||||||||||||||
| 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
-Additional map: Local Refinement of DH-MBD cryoSPARC Density-modified (EMReady)
| File | emd_53972_additional_1.map | ||||||||||||
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| Annotation | Local Refinement of DH-MBD cryoSPARC Density-modified (EMReady) | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: Local Refinement of DH-MBD cryoSPARC Half map 1
| File | emd_53972_half_map_1.map | ||||||||||||
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| Annotation | Local Refinement of DH-MBD cryoSPARC Half map 1 | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: Local Refinement of DH-MBD cryoSPARC Half-map 2
| File | emd_53972_half_map_2.map | ||||||||||||
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| Annotation | Local Refinement of DH-MBD cryoSPARC Half-map 2 | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Phospho-DH bound by Sld3-MBD on ARS1 DNA
+Supramolecule #1: Phospho-DH bound by Sld3-MBD on ARS1 DNA
+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: DNA replication regulator SLD3
+Macromolecule #8: DNA (53-MER)
+Macromolecule #9: DNA (53-MER)
+Macromolecule #10: ZINC ION
+Macromolecule #11: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #12: MAGNESIUM ION
-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: 7.5 |
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| Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 3 / Number real images: 105652 / Average electron dose: 49.8 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: 2.5 µm / Nominal defocus min: 1.1 µm / Nominal magnification: 130000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi




Keywords
Authors
United Kingdom, European Union,
Germany, 6 items
Citation






Z (Sec.)
Y (Row.)
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Processing
FIELD EMISSION GUN

