+
Open data
-
Basic information
| Entry | Database: PDB / ID: 28vy | |||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Title | sCMGE assembled on ARS1 DNA with Sld2 and RPA | |||||||||||||||||||||||||||
Components |
| |||||||||||||||||||||||||||
Keywords | REPLICATION / Macromolecular complex / DNA / ATPase / Helicase MCM2-7 | |||||||||||||||||||||||||||
| 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 | ![]() synthetic construct (others) | |||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.7 Å | |||||||||||||||||||||||||||
Authors | Butryn, A. / Palm, G. / Costa, A. | |||||||||||||||||||||||||||
| Funding support | United Kingdom, European Union, Germany, 4items
| |||||||||||||||||||||||||||
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. | |||||||||||||||||||||||||||
| History |
|
-
Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
|---|
-
Downloads & links
-
Download
| PDBx/mmCIF format | 28vy.cif.gz | 1.7 MB | Display | PDBx/mmCIF format |
|---|---|---|---|---|
| PDB format | pdb28vy.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 28vy.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/8v/28vy ftp://data.pdbj.org/pub/pdb/validation_reports/8v/28vy | HTTPS FTP |
|---|
-Related structure data
| Related structure data | ![]() 9rhiC ![]() 9rhjC ![]() 9rhlC ![]() 9rhmC M: map data used to model this data C: citing same article ( |
|---|---|
| Similar structure data | Similarity search - Function & homology F&H Search |
-
Links
-
Assembly
| Deposited unit | ![]()
|
|---|---|
| 1 |
|
-
Components
-DNA replication licensing factor ... , 5 types, 5 molecules 23467
| #1: Protein | Mass: 98911.539 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: MCM2, YBL023C, YBL0438 / Production host: ![]() |
|---|---|
| #2: Protein | Mass: 111720.242 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: MCM3, YEL032W, SYGP-ORF23 / Production host: ![]() |
| #3: Protein | Mass: 105138.375 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: MCM4, CDC54, HCD21, YPR019W, YP9531.13 / Production host: ![]() |
| #5: Protein | Mass: 113110.211 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: MCM6, YGL201C / Production host: ![]() |
| #6: Protein | Mass: 95049.875 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: MCM7, CDC47, YBR202W, YBR1441 / Production host: ![]() |
-Protein , 2 types, 2 molecules 5E
| #4: Protein | Mass: 86505.734 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: MCM5, CDC46, YLR274W, L9328.1 / Production host: ![]() |
|---|---|
| #10: Protein | Mass: 75154.703 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: CDC45, SLD4, YLR103C, L8004.11 / Production host: ![]() |
-DNA chain , 1 types, 2 molecules AB
| #7: DNA chain | Mass: 8316.371 Da / Num. of mol.: 2 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
|---|
-DNA replication complex GINS protein ... , 4 types, 4 molecules CDHI
| #8: Protein | Mass: 24464.885 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: PSF3, YOL146W / Production host: ![]() |
|---|---|
| #9: Protein | Mass: 33983.617 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: SLD5, YDR489W / Production host: ![]() |
| #13: Protein | Mass: 24230.576 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: PSF1, YDR013W, PZA208, YD8119.18 / Production host: ![]() |
| #14: Protein | Mass: 25096.807 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: PSF2, YJL072C, HRF213, J1086 / Production host: ![]() |
-DNA polymerase epsilon ... , 2 types, 2 molecules FG
| #11: Protein | Mass: 78425.852 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: DPB2, YPR175W, P9705.7 / Production host: ![]() |
|---|---|
| #12: Protein | Mass: 255992.484 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: POL2, DUN2, YNL262W, N0825 / Production host: ![]() References: UniProt: P21951, DNA-directed DNA polymerase, Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters |
-Non-polymers , 4 types, 19 molecules 






| #15: Chemical | ChemComp-MG / #16: Chemical | ChemComp-ZN / #17: Chemical | ChemComp-ADP / | #18: Chemical | ChemComp-ATP / |
|---|
-Details
| Has ligand of interest | N |
|---|---|
| Has protein modification | N |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
|---|---|
| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-
Sample preparation
| Component | Name: Single CMGE complex with RPA on replication origin / Type: COMPLEX / Entity ID: #1-#14 / Source: RECOMBINANT |
|---|---|
| Molecular weight | Experimental value: NO |
| Source (natural) | Organism: ![]() |
| Source (recombinant) | Organism: ![]() |
| Buffer solution | pH: 7.6 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Cryogen name: ETHANE |
-
Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
|---|---|
| Microscopy | Model: TFS KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2900 nm / Nominal defocus min: 2000 nm |
| Image recording | Electron dose: 38.6 e/Å2 / Film or detector model: FEI FALCON IV (4k x 4k) |
-
Processing
| EM software |
| ||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 2.7 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 1125095 / Symmetry type: POINT | ||||||||||||||||||||||||
| Refinement | Cross valid method: NONE Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||
| Displacement parameters | Biso mean: 161.05 Å2 | ||||||||||||||||||||||||
| Refine LS restraints |
|
Movie
Controller
About Yorodumi






United Kingdom, European Union,
Germany, 4items
Citation





PDBj












































FIELD EMISSION GUN