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Yorodumi- PDB-7pmn: S. cerevisiae replisome-SCF(Dia2) complex bound to double-strande... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 7pmn | ||||||||||||
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| Title | S. cerevisiae replisome-SCF(Dia2) complex bound to double-stranded DNA (conformation II) | ||||||||||||
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Keywords | REPLICATION / Genome stability / DNA replication / Ubiquitination / termination / replisome / cryo-EM / CMG / SCF(Dia2) | ||||||||||||
| Function / homology | Function and homology informationRAVE complex / Iron uptake and transport / CBF3 complex / regulation of transcription by galactose / establishment of sister chromatid cohesion / cellular response to methylmercury / vacuolar proton-transporting V-type ATPase complex assembly / mitotic DNA replication termination / gene conversion / : ...RAVE complex / Iron uptake and transport / CBF3 complex / regulation of transcription by galactose / establishment of sister chromatid cohesion / cellular response to methylmercury / vacuolar proton-transporting V-type ATPase complex assembly / mitotic DNA replication termination / gene conversion / : / DNA-templated DNA replication maintenance of fidelity / septin ring assembly / maintenance of DNA repeat elements / Unwinding of DNA / invasive growth in response to glucose limitation / replication fork protection complex / replication fork arrest / DNA replication initiation / meiotic chromosome segregation / protein-containing complex disassembly / 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 / exit from mitosis / 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 / regulation of exit from mitosis / nucleotide-excision repair, DNA gap filling / Activation of the pre-replicative complex / SUMO binding / establishment of mitotic sister chromatid cohesion / nuclear pre-replicative complex / CMG complex / DNA replication proofreading / DNA replication checkpoint signaling / Activation of ATR in response to replication stress / DNA replication preinitiation complex / single-stranded DNA 3'-5' DNA exonuclease activity / positive regulation of D-glucose transmembrane transport / mitotic DNA replication checkpoint signaling / vacuolar acidification / double-strand break repair via break-induced replication / MCM complex / mitotic DNA replication initiation / mitochondrial fusion / mitotic intra-S DNA damage checkpoint signaling / protein neddylation / mitotic DNA replication / silent mating-type cassette heterochromatin formation / single-stranded DNA helicase activity / mitotic sister chromatid cohesion / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / regulation of DNA-templated DNA replication initiation / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / kinetochore assembly / SCF ubiquitin ligase complex / DNA strand elongation involved in DNA replication / leading strand elongation / SCF-dependent proteasomal ubiquitin-dependent protein catabolic process / nuclear chromosome / Antigen processing: Ubiquitination & Proteasome degradation / regulation of DNA replication / nuclear replication fork / DNA replication origin binding / replication fork processing / subtelomeric heterochromatin formation / Dual incision in TC-NER / error-prone translesion synthesis / DNA replication initiation / regulation of protein-containing complex assembly / cullin family protein binding / base-excision repair, gap-filling / negative regulation of cytoplasmic translation / endomembrane system / regulation of mitotic cell cycle / DNA helicase activity / helicase activity / replication fork / G1/S transition of mitotic cell cycle / meiotic cell cycle / transcription elongation by RNA polymerase II / base-excision repair / DNA-templated DNA replication / double-strand break repair via nonhomologous end joining / G2/M transition of mitotic cell cycle / kinetochore / double-strand break repair / mitotic cell cycle / peroxisome / nucleosome assembly / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / heterochromatin formation / protein-containing complex assembly Similarity search - Function | ||||||||||||
| Biological species | ![]() Synthetic construct (others) | ||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||||||||
Authors | Jenkyn-Bedford, M. / Yeeles, J.T.P. / Deegan, T.D. | ||||||||||||
| Funding support | United Kingdom, 3items
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Citation | Journal: Nature / Year: 2021Title: A conserved mechanism for regulating replisome disassembly in eukaryotes. Authors: Michael Jenkyn-Bedford / Morgan L Jones / Yasemin Baris / Karim P M Labib / Giuseppe Cannone / Joseph T P Yeeles / Tom D Deegan / ![]() Abstract: Replisome disassembly is the final step of eukaryotic DNA replication and is triggered by ubiquitylation of the CDC45-MCM-GINS (CMG) replicative helicase. Despite being driven by evolutionarily ...Replisome disassembly is the final step of eukaryotic DNA replication and is triggered by ubiquitylation of the CDC45-MCM-GINS (CMG) replicative helicase. Despite being driven by evolutionarily diverse E3 ubiquitin ligases in different eukaryotes (SCF in budding yeast, CUL2 in metazoa), replisome disassembly is governed by a common regulatory principle, in which ubiquitylation of CMG is suppressed before replication termination, to prevent replication fork collapse. Recent evidence suggests that this suppression is mediated by replication fork DNA. However, it is unknown how SCF and CUL2 discriminate terminated from elongating replisomes, to selectively ubiquitylate CMG only after termination. Here we used cryo-electron microscopy to solve high-resolution structures of budding yeast and human replisome-E3 ligase assemblies. Our structures show that the leucine-rich repeat domains of Dia2 and LRR1 are structurally distinct, but bind to a common site on CMG, including the MCM3 and MCM5 zinc-finger domains. The LRR-MCM interaction is essential for replisome disassembly and, crucially, is occluded by the excluded DNA strand at replication forks, establishing the structural basis for the suppression of CMG ubiquitylation before termination. Our results elucidate a conserved mechanism for the regulation of replisome disassembly in eukaryotes, and reveal a previously unanticipated role for DNA in preserving replisome integrity. | ||||||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7pmn.cif.gz | 1.7 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb7pmn.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 7pmn.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/pm/7pmn ftp://data.pdbj.org/pub/pdb/validation_reports/pm/7pmn | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 13539MC ![]() 7ploC ![]() 7pmkC M: map data used to model this data C: citing same article ( |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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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: ![]() |
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| #2: Protein | Mass: 111987.562 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 , 7 types, 9 molecules 5EFGHKLXY
| #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: ![]() | ||||||||
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| #11: 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: ![]() | ||||||||
| #12: Protein | Mass: 108610.148 Da / Num. of mol.: 3 / Fragment: Mcm6 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: CTF4, CHL15, POB1, YPR135W, P9659.7 / Production host: ![]() #15: Protein | | Mass: 22357.270 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: SKP1, CBF3D, YDR328C, D9798.14 / Production host: ![]() #16: Protein | | Mass: 85368.844 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: DIA2, YOR080W, YOR29-31 / Production host: ![]() #19: Protein | | Mass: 141296.875 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: TOF1, YNL273W, N0636 / Production host: ![]() #20: Protein | | Mass: 36588.758 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: CSM3, YMR048W, YM9796.01 / Production host: ![]() |
-DNA replication complex GINS protein ... , 4 types, 4 molecules ABCD
| #7: Protein | Mass: 24230.576 Da / Num. of mol.: 1 / Fragment: Tof1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: PSF1, YDR013W, PZA208, YD8119.18 / Production host: ![]() |
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| #8: Protein | Mass: 25096.807 Da / Num. of mol.: 1 / Fragment: Csm3 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: PSF2, YJL072C, HRF213, J1086 / Production host: ![]() |
| #9: Protein | Mass: 22004.160 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: PACBIOSEQ_LOCUS5150, PACBIOSEQ_LOCUS5239, PACBIOSEQ_LOCUS5244, PACBIOSEQ_LOCUS5270, PACBIOSEQ_LOCUS5331 Production host: ![]() |
| #10: 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: ![]() |
-DNA chain , 2 types, 2 molecules IJ
| #13: DNA chain | Mass: 37738.676 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Synthetic construct (others) |
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| #14: DNA chain | Mass: 35259.238 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Production host: ![]() |
-DNA polymerase epsilon ... , 2 types, 2 molecules QR
| #17: Protein | Mass: 255890.422 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 |
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| #18: 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: ![]() |
-Non-polymers , 3 types, 11 molecules 




| #21: Chemical | | #22: Chemical | #23: Chemical | ChemComp-ZN / |
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-Details
| Has ligand of interest | Y |
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-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
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| Molecular weight | Experimental value: NO | ||||||||||||||||||||||||||||
| Source (natural) |
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| Source (recombinant) |
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| Buffer solution | pH: 7.6 | ||||||||||||||||||||||||||||
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||
| Specimen support | Details: 15 mA / Grid material: COPPER / Grid mesh size: 400 divisions/in. / Grid type: Quantifoil R2/2 | ||||||||||||||||||||||||||||
| Vitrification | Cryogen name: ETHANE / Details: Manual plunger |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TITAN KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 81000 X / Nominal defocus max: 2200 nm / Nominal defocus min: 400 nm / Alignment procedure: COMA FREE |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Average exposure time: 4 sec. / Electron dose: 38.8 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of real images: 12730 |
| EM imaging optics | Energyfilter name: GIF Bioquantum / Energyfilter slit width: 20 eV |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 2160000 | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 369254 Details: Composite map produced from individual cryo-EM density maps (resolutions 3.2 - 4.0 A) using Phenix combine_focused_maps. See publication for details. Symmetry type: POINT |
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