+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10930 | |||||||||
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Title | Cohesin complex with loader gripping DNA | |||||||||
Map data | ||||||||||
Sample |
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Keywords | Chromosome segregation Sister chromatid cohesion SMC complexes Cohesin ABC-ATPase Mis4-Scc2-NIPBL / DNA BINDING PROTEIN | |||||||||
Function / homology | Function and homology information pericentric heterochromatin => GO:0005721 / meiotic cohesin complex => GO:0030893 / mitotic cohesin dsDNA (leading strand) loading / positive regulation of mitotic cohesin loading / Cohesin Loading onto Chromatin / Resolution of Sister Chromatid Cohesion / mitotic cohesin ssDNA (lagging strand) loading / maintenance of mitotic sister chromatid cohesion, centromeric / mitotic cohesin complex => GO:0030892 / : ...pericentric heterochromatin => GO:0005721 / meiotic cohesin complex => GO:0030893 / mitotic cohesin dsDNA (leading strand) loading / positive regulation of mitotic cohesin loading / Cohesin Loading onto Chromatin / Resolution of Sister Chromatid Cohesion / mitotic cohesin ssDNA (lagging strand) loading / maintenance of mitotic sister chromatid cohesion, centromeric / mitotic cohesin complex => GO:0030892 / : / SUMOylation of DNA damage response and repair proteins / nucleus leading edge / SMC loading complex / mitotic telomere tethering at nuclear periphery / Scc2-Scc4 cohesin loading complex / mitotic cohesin loading / cohesin loader activity / heterochromatin island / maintenance of mitotic sister chromatid cohesion / cohesin complex / mitotic cohesin complex / chromosome, subtelomeric region / establishment of protein localization to chromatin / rDNA heterochromatin / replication-born double-strand break repair via sister chromatid exchange / establishment of mitotic sister chromatid cohesion / condensed chromosome, centromeric region / sister chromatid cohesion / mitotic sister chromatid cohesion / nuclear chromosome / mitotic sister chromatid segregation / pericentric heterochromatin / enzyme regulator activity / chromosome / double-strand break repair / double-stranded DNA binding / single-stranded DNA binding / cell division / chromatin binding / DNA damage response / chromatin / DNA binding / ATP binding / nucleus / cytosol Similarity search - Function | |||||||||
Biological species | Schizosaccharomyces pombe (strain 972 / ATCC 24843) (yeast) / Synthetic construct (others) / synthetic construct (others) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.94 Å | |||||||||
Authors | Higashi TL / Eickhoff P | |||||||||
Funding support | 2 items
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Citation | Journal: Mol Cell / Year: 2020 Title: A Structure-Based Mechanism for DNA Entry into the Cohesin Ring. Authors: Torahiko L Higashi / Patrik Eickhoff / Joana S Sousa / Julia Locke / Andrea Nans / Helen R Flynn / Ambrosius P Snijders / George Papageorgiou / Nicola O'Reilly / Zhuo A Chen / Francis J ...Authors: Torahiko L Higashi / Patrik Eickhoff / Joana S Sousa / Julia Locke / Andrea Nans / Helen R Flynn / Ambrosius P Snijders / George Papageorgiou / Nicola O'Reilly / Zhuo A Chen / Francis J O'Reilly / Juri Rappsilber / Alessandro Costa / Frank Uhlmann / Abstract: Despite key roles in sister chromatid cohesion and chromosome organization, the mechanism by which cohesin rings are loaded onto DNA is still unknown. Here we combine biochemical approaches and ...Despite key roles in sister chromatid cohesion and chromosome organization, the mechanism by which cohesin rings are loaded onto DNA is still unknown. Here we combine biochemical approaches and cryoelectron microscopy (cryo-EM) to visualize a cohesin loading intermediate in which DNA is locked between two gates that lead into the cohesin ring. Building on this structural framework, we design experiments to establish the order of events during cohesin loading. In an initial step, DNA traverses an N-terminal kleisin gate that is first opened upon ATP binding and then closed as the cohesin loader locks the DNA against the ATPase gate. ATP hydrolysis will lead to ATPase gate opening to complete DNA entry. Whether DNA loading is successful or results in loop extrusion might be dictated by a conserved kleisin N-terminal tail that guides the DNA through the kleisin gate. Our results establish the molecular basis for cohesin loading onto DNA. | |||||||||
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_10930.map.gz | 167.7 MB | EMDB map data format | |
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Header (meta data) | emd-10930-v30.xml emd-10930.xml | 28.2 KB 28.2 KB | Display Display | EMDB header |
Images | emd_10930.png | 174.6 KB | ||
Filedesc metadata | emd-10930.cif.gz | 9.2 KB | ||
Others | emd_10930_additional.map.gz emd_10930_half_map_1.map.gz emd_10930_half_map_2.map.gz | 11.5 MB 165 MB 165 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10930 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10930 | HTTPS FTP |
-Validation report
Summary document | emd_10930_validation.pdf.gz | 938.7 KB | Display | EMDB validaton report |
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Full document | emd_10930_full_validation.pdf.gz | 938.3 KB | Display | |
Data in XML | emd_10930_validation.xml.gz | 14.9 KB | Display | |
Data in CIF | emd_10930_validation.cif.gz | 17.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10930 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10930 | HTTPS FTP |
-Related structure data
Related structure data | 6yufMC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_10930.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Voxel size | X=Y=Z: 1.09 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: density modified map produced with Phenix resolve cryo-EM tool
File | emd_10930_additional.map | ||||||||||||
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Annotation | density modified map produced with Phenix resolve cryo-EM tool | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_10930_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_10930_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Cohesin complex with loader gripping DNA
+Supramolecule #1: Cohesin complex with loader gripping DNA
+Supramolecule #2: Cohesin subunit rad21, Structural maintenance of chromosomes prot...
+Supramolecule #3: DNA (32-MER)
+Supramolecule #4: Sister chromatid cohesion protein mis4
+Macromolecule #1: Cohesin subunit rad21
+Macromolecule #2: Sister chromatid cohesion protein mis4
+Macromolecule #3: Structural maintenance of chromosomes protein 1
+Macromolecule #4: Structural maintenance of chromosomes protein 3
+Macromolecule #5: DNA (32-MER)
+Macromolecule #6: DNA (32-MER)
+Macromolecule #7: BERYLLIUM TRIFLUORIDE ION
+Macromolecule #8: 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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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Average electron dose: 33.8 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |