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Open data
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Basic information
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| Title | Cryo-EM structure of the human MCM2-7 heterohexamer | ||||||||||||
Map data | unsharpened map | ||||||||||||
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Keywords | complex / helicase / replication / AAA+ ATPase / DNA BINDING PROTEIN | ||||||||||||
| Function / homology | Function and homology informationSwitching of origins to a post-replicative state / Unwinding of DNA / nuclear origin of replication recognition complex / alpha DNA polymerase:primase complex / Regulation of MITF-M-dependent genes involved in DNA replication, damage repair and senescence / mitotic DNA replication / regulation of phosphorylation / CMG complex / MCM complex / double-strand break repair via break-induced replication ...Switching of origins to a post-replicative state / Unwinding of DNA / nuclear origin of replication recognition complex / alpha DNA polymerase:primase complex / Regulation of MITF-M-dependent genes involved in DNA replication, damage repair and senescence / mitotic DNA replication / regulation of phosphorylation / CMG complex / MCM complex / 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 / DNA replication origin binding / cochlea development / DNA replication initiation / Activation of the pre-replicative complex / Activation of ATR in response to replication stress / cellular response to interleukin-4 / DNA helicase activity / cellular response to epidermal growth factor stimulus / Assembly of the pre-replicative complex / helicase activity / cellular response to xenobiotic stimulus / Orc1 removal from chromatin / nucleosome assembly / single-stranded DNA binding / histone binding / DNA helicase / chromosome, telomeric region / DNA replication / cell population proliferation / cilium / intracellular membrane-bounded organelle / apoptotic process / DNA damage response / chromatin / perinuclear region of cytoplasm / enzyme binding / ATP hydrolysis activity / DNA binding / zinc ion binding / nucleoplasm / ATP binding / identical protein binding / nucleus / membrane / cytoplasm / cytosol Similarity search - Function | ||||||||||||
| Biological species | Homo sapiens (human) | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.5 Å | ||||||||||||
Authors | Yang R / Hunker O / Bleichert F | ||||||||||||
| Funding support | United States, 3 items
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Citation | Journal: Nature / Year: 2024Title: Multiple mechanisms for licensing human replication origins. Authors: Ran Yang / Olivia Hunker / Marleigh Wise / Franziska Bleichert / ![]() Abstract: Loading of replicative helicases is obligatory for the assembly of DNA replication machineries. The eukaryotic MCM2-7 replicative helicase motor is deposited onto DNA by the origin recognition ...Loading of replicative helicases is obligatory for the assembly of DNA replication machineries. The eukaryotic MCM2-7 replicative helicase motor is deposited onto DNA by the origin recognition complex (ORC) and co-loader proteins as a head-to-head double hexamer to license replication origins. Although extensively studied in budding yeast, the mechanisms of origin licensing in multicellular eukaryotes remain poorly defined. Here we use biochemical reconstitution and electron microscopy to reconstruct the human MCM loading pathway. We find that unlike in yeast, the ORC6 subunit of the ORC is not essential for-but enhances-human MCM loading. Electron microscopy analyses identify several intermediates en route to MCM double hexamer formation in the presence and absence of ORC6, including a DNA-loaded, closed-ring MCM single hexamer intermediate that can mature into a head-to-head double hexamer through multiple mechanisms. ORC6 and ORC3 facilitate the recruitment of the ORC to the dimerization interface of the first hexamer into MCM-ORC (MO) complexes that are distinct from the yeast MO complex and may orient the ORC for second MCM hexamer loading. Additionally, MCM double hexamer formation can proceed through dimerization of independently loaded MCM single hexamers, promoted by a propensity of human MCM2-7 hexamers to self-dimerize. This flexibility in human MCM loading may provide resilience against cellular replication stress, and the reconstitution system will enable studies addressing outstanding questions regarding DNA replication initiation and replication-coupled events in the future. | ||||||||||||
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Structure visualization
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_43710.map.gz | 408.3 MB | EMDB map data format | |
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| Header (meta data) | emd-43710-v30.xml emd-43710.xml | 27.8 KB 27.8 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_43710_fsc.xml | 19.9 KB | Display | FSC data file |
| Images | emd_43710.png | 108.4 KB | ||
| Filedesc metadata | emd-43710.cif.gz | 9 KB | ||
| Others | emd_43710_additional_1.map.gz emd_43710_half_map_1.map.gz emd_43710_half_map_2.map.gz | 679 MB 764.6 MB 764.6 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-43710 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-43710 | HTTPS FTP |
-Validation report
| Summary document | emd_43710_validation.pdf.gz | 989.8 KB | Display | EMDB validaton report |
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| Full document | emd_43710_full_validation.pdf.gz | 989.4 KB | Display | |
| Data in XML | emd_43710_validation.xml.gz | 27.6 KB | Display | |
| Data in CIF | emd_43710_validation.cif.gz | 35.9 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43710 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43710 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 8w0iMC ![]() 8w0eC ![]() 8w0fC ![]() 8w0gC ![]() 9caqC C: citing same article ( M: atomic model generated by this map |
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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_43710.map.gz / Format: CCP4 / Size: 824 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | unsharpened map | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.832 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: deepEMhancer map
| File | emd_43710_additional_1.map | ||||||||||||
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| Annotation | deepEMhancer map | ||||||||||||
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| Density Histograms |
-Half map: half-map
| File | emd_43710_half_map_1.map | ||||||||||||
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| Annotation | half-map | ||||||||||||
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| Density Histograms |
-Half map: half-map
| File | emd_43710_half_map_2.map | ||||||||||||
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| Annotation | half-map | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : human MCM2-7 heterohexamer
+Supramolecule #1: human MCM2-7 heterohexamer
+Macromolecule #1: DNA replication licensing factor MCM2
+Macromolecule #2: DNA replication licensing factor MCM3
+Macromolecule #3: DNA replication licensing factor MCM4
+Macromolecule #4: DNA replication licensing factor MCM5
+Macromolecule #5: DNA replication licensing factor MCM6
+Macromolecule #6: DNA replication licensing factor MCM7
+Macromolecule #7: ZINC ION
+Macromolecule #8: MAGNESIUM ION
+Macromolecule #9: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #10: ADENOSINE-5'-DIPHOSPHATE
-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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| Grid | Model: Quantifoil R2/1 / Material: COPPER / Mesh: 300 / Support film - Material: GRAPHENE OXIDE |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.96 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi




Keywords
Homo sapiens (human)
Authors
United States, 3 items
Citation











Z (Sec.)
Y (Row.)
X (Col.)












































Trichoplusia ni (cabbage looper)

Processing
FIELD EMISSION GUN

