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Yorodumi- EMDB-13534: Cryo-EM map of the core human replisome on a DNA substrate lackin... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13534 | ||||||||||||
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Title | Cryo-EM map of the core human replisome on a DNA substrate lacking a 5'-flap | ||||||||||||
Map data | Core human replisome on fork DNA lacking a 5'-flap | ||||||||||||
Sample |
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Function / homology | Function and homology information cellular response to bleomycin / DNA secondary structure binding / detection of abiotic stimulus / replication fork arrest / regulation of nuclear cell cycle DNA replication / Switching of origins to a post-replicative state / Unwinding of DNA / cell cycle phase transition / cellular response to cisplatin / DNA replication initiation ...cellular response to bleomycin / DNA secondary structure binding / detection of abiotic stimulus / replication fork arrest / regulation of nuclear cell cycle DNA replication / Switching of origins to a post-replicative state / Unwinding of DNA / cell cycle phase transition / cellular response to cisplatin / DNA replication initiation / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / GINS complex / mitotic DNA replication preinitiation complex assembly / nuclear origin of replication recognition complex / cellular response to hydroxyurea / anaphase-promoting complex binding / cullin-RING-type E3 NEDD8 transferase / alpha DNA polymerase:primase complex / mitotic DNA replication / cellular response to chemical stress / NEDD8 transferase activity / cullin-RING ubiquitin ligase complex / CMG complex / DNA replication checkpoint signaling / ubiquitin-dependent protein catabolic process via the C-end degron rule pathway / Cul7-RING ubiquitin ligase complex / single-stranded DNA 3'-5' DNA exonuclease activity / DNA replication preinitiation complex / nucleotide-excision repair, DNA gap filling / target-directed miRNA degradation / Loss of Function of FBXW7 in Cancer and NOTCH1 Signaling / regulation of phosphorylation / MCM complex / elongin complex / VCB complex / DNA replication proofreading / mitotic DNA replication checkpoint signaling / replication fork protection complex / positive regulation of protein autoubiquitination / double-strand break repair via break-induced replication / protein neddylation / mitotic DNA replication initiation / mitotic intra-S DNA damage checkpoint signaling / NEDD8 ligase activity / positive regulation of double-strand break repair / Cul5-RING ubiquitin ligase complex / negative regulation of response to oxidative stress / regulation of DNA-templated DNA replication initiation / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / inner cell mass cell proliferation / ubiquitin-ubiquitin ligase activity / SCF ubiquitin ligase complex / Cul2-RING ubiquitin ligase complex / Cul4A-RING E3 ubiquitin ligase complex / negative regulation of type I interferon production / ubiquitin ligase complex scaffold activity / SCF-dependent proteasomal ubiquitin-dependent protein catabolic process / Cul4B-RING E3 ubiquitin ligase complex / Cul3-RING ubiquitin ligase complex / DNA strand elongation involved in DNA replication / branching morphogenesis of an epithelial tube / activation of protein kinase activity / DNA synthesis involved in DNA repair / cochlea development / G1/S-Specific Transcription / DNA unwinding involved in DNA replication / leading strand elongation / Apoptotic cleavage of cellular proteins / replication fork processing / Prolactin receptor signaling / nuclear replication fork / protein monoubiquitination / mitotic G2 DNA damage checkpoint signaling / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / cullin family protein binding / DNA replication origin binding / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / positive regulation of double-strand break repair via homologous recombination / PCNA-Dependent Long Patch Base Excision Repair / Activation of the pre-replicative complex / Tat-mediated elongation of the HIV-1 transcript / DNA replication initiation / Formation of HIV-1 elongation complex containing HIV-1 Tat / embryonic organ development / cellular response to interleukin-4 / Formation of HIV elongation complex in the absence of HIV Tat / protein K48-linked ubiquitination / error-prone translesion synthesis / Nuclear events stimulated by ALK signaling in cancer / Activation of ATR in response to replication stress / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / response to UV / RNA Polymerase II Pre-transcription Events / base-excision repair, gap-filling / T cell activation / positive regulation of TORC1 signaling Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | ||||||||||||
Authors | Jones MJ / Yeeles JTP / Jenkyn-Bedford M / Deegan TD | ||||||||||||
Funding support | United Kingdom, 3 items
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Citation | Journal: Nature / Year: 2021 Title: 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. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_13534.map.gz | 192.9 MB | EMDB map data format | |
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Header (meta data) | emd-13534-v30.xml emd-13534.xml | 14.1 KB 14.1 KB | Display Display | EMDB header |
Images | emd_13534.png | 77.8 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13534 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13534 | HTTPS FTP |
-Validation report
Summary document | emd_13534_validation.pdf.gz | 333.2 KB | Display | EMDB validaton report |
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Full document | emd_13534_full_validation.pdf.gz | 332.8 KB | Display | |
Data in XML | emd_13534_validation.xml.gz | 7.1 KB | Display | |
Data in CIF | emd_13534_validation.cif.gz | 8.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13534 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13534 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_13534.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Core human replisome on fork DNA lacking a 5'-flap | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.072 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Core human replisome lacking a 5'-flap
Entire | Name: Core human replisome lacking a 5'-flap |
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Components |
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-Supramolecule #1: Core human replisome lacking a 5'-flap
Supramolecule | Name: Core human replisome lacking a 5'-flap / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#24 |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.6 |
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 16721 / Average exposure time: 4.0 sec. / Average electron dose: 38.8 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 0.1 mm / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 0.8 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |