+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-25924 | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | S. cerevisiae ORC bound to 84 bp ARS1 DNA | ||||||||||||
Map data | unsharpened cryo-EM map | ||||||||||||
Sample |
| ||||||||||||
Keywords | initiator / helicase loader / AAA+ ATPase / REPLICATION-DNA complex | ||||||||||||
Function / homology | Function and homology information CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / Assembly of the ORC complex at the origin of replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear origin of replication recognition complex / Activation of the pre-replicative complex / nucleosome organization / nuclear pre-replicative complex / Activation of ATR in response to replication stress ...CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / Assembly of the ORC complex at the origin of replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear origin of replication recognition complex / Activation of the pre-replicative complex / nucleosome organization / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / mitotic DNA replication checkpoint signaling / silent mating-type cassette heterochromatin formation / Orc1 removal from chromatin / regulation of DNA replication / DNA replication origin binding / DNA replication initiation / subtelomeric heterochromatin formation / nucleosome binding / chromosome, telomeric region / chromatin binding / ATP hydrolysis activity / nucleoplasm / ATP binding / nucleus / metal ion binding Similarity search - Function | ||||||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.6 Å | ||||||||||||
Authors | Schmidt JM / Yang R | ||||||||||||
Funding support | United States, 3 items
| ||||||||||||
Citation | Journal: Nat Commun / Year: 2022 Title: A mechanism of origin licensing control through autoinhibition of S. cerevisiae ORC·DNA·Cdc6. Authors: Jan Marten Schmidt / Ran Yang / Ashish Kumar / Olivia Hunker / Jan Seebacher / Franziska Bleichert / Abstract: The coordinated action of multiple replicative helicase loading factors is needed for the licensing of replication origins prior to DNA replication. Binding of the Origin Recognition Complex (ORC) to ...The coordinated action of multiple replicative helicase loading factors is needed for the licensing of replication origins prior to DNA replication. Binding of the Origin Recognition Complex (ORC) to DNA initiates the ATP-dependent recruitment of Cdc6, Cdt1 and Mcm2-7 loading, but the structural details for timely ATPase site regulation and for how loading can be impeded by inhibitory signals, such as cyclin-dependent kinase phosphorylation, are unknown. Using cryo-electron microscopy, we have determined several structures of S. cerevisiae ORC·DNA·Cdc6 intermediates at 2.5-2.7 Å resolution. These structures reveal distinct ring conformations of the initiator·co-loader assembly and inactive ATPase site configurations for ORC and Cdc6. The Orc6 N-terminal domain laterally engages the ORC·Cdc6 ring in a manner that is incompatible with productive Mcm2-7 docking, while deletion of this Orc6 region alleviates the CDK-mediated inhibition of Mcm7 recruitment. Our findings support a model in which Orc6 promotes the assembly of an autoinhibited ORC·DNA·Cdc6 intermediate to block origin licensing in response to CDK phosphorylation and to avert DNA re-replication. | ||||||||||||
History |
|
-Structure visualization
Supplemental images |
---|
-Downloads & links
-EMDB archive
Map data | emd_25924.map.gz | 80.6 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-25924-v30.xml emd-25924.xml | 30.8 KB 30.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_25924_fsc.xml | 10.7 KB | Display | FSC data file |
Images | emd_25924.png | 108.5 KB | ||
Filedesc metadata | emd-25924.cif.gz | 8.6 KB | ||
Others | emd_25924_additional_1.map.gz emd_25924_additional_2.map.gz emd_25924_half_map_1.map.gz emd_25924_half_map_2.map.gz | 9.7 MB 13.9 MB 80.8 MB 80.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-25924 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25924 | HTTPS FTP |
-Validation report
Summary document | emd_25924_validation.pdf.gz | 934.1 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_25924_full_validation.pdf.gz | 933.7 KB | Display | |
Data in XML | emd_25924_validation.xml.gz | 17.5 KB | Display | |
Data in CIF | emd_25924_validation.cif.gz | 23.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25924 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25924 | HTTPS FTP |
-Related structure data
Related structure data | 7tjfMC 7tjhC 7tjiC 7tjjC 7tjkC C: citing same article (ref.) M: atomic model generated by this map |
---|---|
Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_25924.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | unsharpened cryo-EM map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.86 Å | ||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
|
-Supplemental data
-Additional map: sharpened, masked cryo-EM map
File | emd_25924_additional_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | sharpened, masked cryo-EM map | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Additional map: density modified cryo-EM map
File | emd_25924_additional_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | density modified cryo-EM map | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: half map
File | emd_25924_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | half map | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: half map
File | emd_25924_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | half map | ||||||||||||
Projections & Slices |
| ||||||||||||
Density Histograms |
-Sample components
+Entire : ORC in complex with ARS1 DNA
+Supramolecule #1: ORC in complex with ARS1 DNA
+Macromolecule #1: Origin recognition complex subunit 1
+Macromolecule #2: Origin recognition complex subunit 2
+Macromolecule #3: Origin recognition complex subunit 3
+Macromolecule #4: Origin recognition complex subunit 4
+Macromolecule #5: Origin recognition complex subunit 5
+Macromolecule #6: Origin recognition complex subunit 6
+Macromolecule #7: DNA, 84 bp ARS1
+Macromolecule #8: DNA, 84 bp ARS1
+Macromolecule #9: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #10: MAGNESIUM ION
+Macromolecule #11: water
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.6 |
---|---|
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 6.6 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: 1.6 µm / Nominal defocus min: 0.8 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |