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Open data
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Basic information
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| Title | pre-Initiation Complex on ARS1 DNA (monomer) | |||||||||||||||
Map data | Local Refinement of pre-IC monomer cryoSPARC Unsharpened map | |||||||||||||||
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Keywords | Macromolecular Complex DNA ATPase Helicase MCM2-7 / REPLICATION | |||||||||||||||
| Function / homology | Function and homology informationregulation of mitotic DNA replication initiation / mating type switching / 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 ...regulation of mitotic DNA replication initiation / mating type switching / 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 / Activation of the pre-replicative complex / mitotic DNA replication / DNA replication checkpoint signaling / 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 / recombinational repair / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / mitotic G2 DNA damage checkpoint signaling / nuclear replication fork / DNA replication origin binding / Dual incision in TC-NER / chromosome, centromeric region / error-prone translesion synthesis / DNA replication initiation / protein kinase activator activity / subtelomeric heterochromatin formation / DNA polymerase binding / replication fork / DNA helicase activity / helicase activity / transcription elongation by RNA polymerase II / DNA-templated DNA replication / spindle pole / nuclear envelope / peroxisome / heterochromatin formation / single-stranded DNA binding / DNA helicase / DNA replication / chromosome, telomeric region / chromatin binding / DNA damage response / chromatin / endoplasmic reticulum / 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: 3.2 Å | |||||||||||||||
Authors | Puehringer T / Butryn A / Couves EC / 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_53970.map.gz | 411 MB | EMDB map data format | |
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| Header (meta data) | emd-53970-v30.xml emd-53970.xml | 51.4 KB 51.4 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_53970_fsc.xml | 19.8 KB | Display | FSC data file |
| Images | emd_53970.png | 51.6 KB | ||
| Filedesc metadata | emd-53970.cif.gz | 14 KB | ||
| Others | emd_53970_additional_1.map.gz emd_53970_half_map_1.map.gz emd_53970_half_map_2.map.gz | 479.4 MB 764.8 MB 764.8 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-53970 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-53970 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9rhiMC ![]() 28vyC ![]() 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_53970.map.gz / Format: CCP4 / Size: 824 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Local Refinement of pre-IC monomer cryoSPARC Unsharpened map | ||||||||||||||||||||||||||||||||||||
| 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: Local Refinement of pre-IC monomer cryoSPARC Density-modified (EMReady)...
| File | emd_53970_additional_1.map | ||||||||||||
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| Annotation | Local Refinement of pre-IC monomer cryoSPARC Density-modified (EMReady) | ||||||||||||
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| Density Histograms |
-Half map: Local Refinement of pre-IC monomer cryoSPARC Half map 1
| File | emd_53970_half_map_1.map | ||||||||||||
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| Annotation | Local Refinement of pre-IC monomer cryoSPARC Half map 1 | ||||||||||||
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| Density Histograms |
-Half map: Local Refinement of pre-IC monomer cryoSPARC Half map 2
| File | emd_53970_half_map_2.map | ||||||||||||
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| Annotation | Local Refinement of pre-IC monomer cryoSPARC Half map 2 | ||||||||||||
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| Density Histograms |
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Sample components
+Entire : Pre-initiation complex (Monomer) on replication origin (DNA)
+Supramolecule #1: Pre-initiation complex (Monomer) on replication origin (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 #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 replication complex GINS protein PSF1
+Macromolecule #14: DNA replication complex GINS protein PSF2
+Macromolecule #15: DNA replication regulator SLD3
+Macromolecule #16: Mitochondrial morphogenesis protein SLD7
+Macromolecule #17: DNA replication regulator DPB11
+Macromolecule #7: DNA (31-MER)
+Macromolecule #8: DNA (31-MER)
+Macromolecule #18: ZINC ION
+Macromolecule #19: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #20: 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: FEI FALCON IV (4k x 4k) / Number grids imaged: 2 / Number real images: 59347 / Average electron dose: 39.0 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: 3.0 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
Citation












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Processing
FIELD EMISSION GUN

