+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-19619 | |||||||||||||||
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Title | H. sapiens MCM2-7 hexamer bound to double stranded DNA | |||||||||||||||
Map data | Map of the human MCM2-7 hexamer bound to double stranded DNA. The map is the output of a local refinement in cryoSPARC. | |||||||||||||||
Sample |
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Keywords | AAA+ ATPase / DNA helicase / REPLICATION | |||||||||||||||
Function / homology | Function and homology information Switching of origins to a post-replicative state / Unwinding of DNA / nuclear origin of replication recognition complex / mitotic DNA replication / alpha DNA polymerase:primase complex / CMG complex / MCM complex / regulation of phosphorylation / double-strand break repair via break-induced replication / mitotic DNA replication initiation ...Switching of origins to a post-replicative state / Unwinding of DNA / nuclear origin of replication recognition complex / mitotic DNA replication / alpha DNA polymerase:primase complex / CMG complex / MCM complex / regulation of phosphorylation / double-strand break repair via break-induced replication / mitotic DNA replication initiation / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / cochlea development / DNA unwinding involved in DNA replication / DNA replication origin binding / DNA replication initiation / Activation of the pre-replicative complex / cellular response to interleukin-4 / Activation of ATR in response to replication stress / DNA helicase activity / Assembly of the pre-replicative complex / helicase activity / Orc1 removal from chromatin / cellular response to xenobiotic stimulus / nucleosome assembly / single-stranded DNA binding / histone binding / DNA helicase / DNA replication / cell population proliferation / chromosome, telomeric region / centrosome / DNA damage response / chromatin / apoptotic process / perinuclear region of cytoplasm / enzyme binding / ATP hydrolysis activity / DNA binding / nucleoplasm / ATP binding / identical protein binding / membrane / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | synthetic construct (others) / Homo sapiens (human) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||||||||
Authors | Greiwe JF / Weissmann F / Diffley JFX / Costa A | |||||||||||||||
Funding support | United Kingdom, European Union, 4 items
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Citation | Journal: Acta Crystallogr D Struct Biol / Year: 2019 Title: Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Authors: Dorothee Liebschner / Pavel V Afonine / Matthew L Baker / Gábor Bunkóczi / Vincent B Chen / Tristan I Croll / Bradley Hintze / Li Wei Hung / Swati Jain / Airlie J McCoy / Nigel W Moriarty ...Authors: Dorothee Liebschner / Pavel V Afonine / Matthew L Baker / Gábor Bunkóczi / Vincent B Chen / Tristan I Croll / Bradley Hintze / Li Wei Hung / Swati Jain / Airlie J McCoy / Nigel W Moriarty / Robert D Oeffner / Billy K Poon / Michael G Prisant / Randy J Read / Jane S Richardson / David C Richardson / Massimo D Sammito / Oleg V Sobolev / Duncan H Stockwell / Thomas C Terwilliger / Alexandre G Urzhumtsev / Lizbeth L Videau / Christopher J Williams / Paul D Adams / Abstract: Diffraction (X-ray, neutron and electron) and electron cryo-microscopy are powerful methods to determine three-dimensional macromolecular structures, which are required to understand biological ...Diffraction (X-ray, neutron and electron) and electron cryo-microscopy are powerful methods to determine three-dimensional macromolecular structures, which are required to understand biological processes and to develop new therapeutics against diseases. The overall structure-solution workflow is similar for these techniques, but nuances exist because the properties of the reduced experimental data are different. Software tools for structure determination should therefore be tailored for each method. Phenix is a comprehensive software package for macromolecular structure determination that handles data from any of these techniques. Tasks performed with Phenix include data-quality assessment, map improvement, model building, the validation/rebuilding/refinement cycle and deposition. Each tool caters to the type of experimental data. The design of Phenix emphasizes the automation of procedures, where possible, to minimize repetitive and time-consuming manual tasks, while default parameters are chosen to encourage best practice. A graphical user interface provides access to many command-line features of Phenix and streamlines the transition between programs, project tracking and re-running of previous tasks. | |||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_19619.map.gz | 229.8 MB | EMDB map data format | |
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Header (meta data) | emd-19619-v30.xml emd-19619.xml | 36.8 KB 36.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_19619_fsc.xml | 13.2 KB | Display | FSC data file |
Images | emd_19619.png | 162.7 KB | ||
Filedesc metadata | emd-19619.cif.gz | 10.8 KB | ||
Others | emd_19619_half_map_1.map.gz emd_19619_half_map_2.map.gz | 226.2 MB 226.2 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-19619 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-19619 | HTTPS FTP |
-Validation report
Summary document | emd_19619_validation.pdf.gz | 1.3 MB | Display | EMDB validaton report |
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Full document | emd_19619_full_validation.pdf.gz | 1.3 MB | Display | |
Data in XML | emd_19619_validation.xml.gz | 22.2 KB | Display | |
Data in CIF | emd_19619_validation.cif.gz | 28.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-19619 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-19619 | HTTPS FTP |
-Related structure data
Related structure data | 8s0aMC 8s09C 8s0bC 8s0cC 8s0dC 8s0eC 8s0fC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_19619.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Map of the human MCM2-7 hexamer bound to double stranded DNA. The map is the output of a local refinement in cryoSPARC. | ||||||||||||||||||||||||||||||||||||
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
-Half map: Map of the human MCM2-7 hexamer bound to...
File | emd_19619_half_map_1.map | ||||||||||||
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Annotation | Map of the human MCM2-7 hexamer bound to double stranded DNA. The map is the output of a local refinement in cryoSPARC. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 1 of the main map.
File | emd_19619_half_map_2.map | ||||||||||||
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Annotation | Half map 1 of the main map. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : H. sapiens MCM2-7 hexamer bound to double stranded DNA
+Supramolecule #1: H. sapiens MCM2-7 hexamer bound to double stranded DNA
+Supramolecule #2: Double stranded DNA
+Supramolecule #3: H. sapiens MCM2-7
+Macromolecule #1: DNA (22-mer)
+Macromolecule #2: DNA (22-mer)
+Macromolecule #3: DNA replication licensing factor MCM2
+Macromolecule #4: DNA replication licensing factor MCM3
+Macromolecule #5: DNA replication licensing factor MCM4
+Macromolecule #6: DNA replication licensing factor MCM5
+Macromolecule #7: DNA replication licensing factor MCM6
+Macromolecule #8: DNA replication licensing factor MCM7
+Macromolecule #9: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #10: MAGNESIUM ION
+Macromolecule #11: ZINC ION
+Macromolecule #12: ADENOSINE-5'-DIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GRAPHENE OXIDE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV |
Details | The origin licensing reaction was reconstituted in vitro using purified H. sapiens proteins, a short DNA template and ATP. Four microlitres of the entire reaction was applied on a grid and incubated for 1 min at room temperature before blotting with filter paper for 5 s and plunge-freezing in liquid ethane. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 3589 / Average exposure time: 9.4 sec. / Average electron dose: 49.19 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: 1.0 µm / Nominal magnification: 130000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model | Chain - Source name: Other / Chain - Initial model type: in silico model / Details: AlphaFold multimer |
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Output model | PDB-8s0a: |