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Yorodumi- EMDB-1835: The lockwasher state of the Drosophila melanogaster Mcm2-7 complex -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-1835 | |||||||||
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Title | The lockwasher state of the Drosophila melanogaster Mcm2-7 complex | |||||||||
Map data | Lockwasher configuration of the Drosophila melanogaster Mcm2-7 | |||||||||
Sample |
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Keywords | DNA replication / Helicase / AAA+ ATPase | |||||||||
Biological species | Drosophila melanogaster (fruit fly) | |||||||||
Method | single particle reconstruction / Resolution: 35.0 Å | |||||||||
Authors | Costa A / Ilves I / Tamberg N / Petojevic T / Nogales E / Botchan MR / Berger JM | |||||||||
Citation | Journal: Nat Struct Mol Biol / Year: 2011 Title: The structural basis for MCM2-7 helicase activation by GINS and Cdc45. Authors: Alessandro Costa / Ivar Ilves / Nele Tamberg / Tatjana Petojevic / Eva Nogales / Michael R Botchan / James M Berger / Abstract: Two central steps for initiating eukaryotic DNA replication involve loading of the Mcm2-7 helicase onto double-stranded DNA and its activation by GINS-Cdc45. To better understand these events, we ...Two central steps for initiating eukaryotic DNA replication involve loading of the Mcm2-7 helicase onto double-stranded DNA and its activation by GINS-Cdc45. To better understand these events, we determined the structures of Mcm2-7 and the CMG complex by using single-particle electron microscopy. Mcm2-7 adopts two conformations--a lock-washer-shaped spiral state and a planar, gapped-ring form--in which Mcm2 and Mcm5 flank a breach in the helicase perimeter. GINS and Cdc45 bridge this gap, forming a topologically closed assembly with a large interior channel; nucleotide binding further seals off the discontinuity between Mcm2 and Mcm5, partitioning the channel into two smaller pores. Together, our data help explain how GINS and Cdc45 activate Mcm2-7, indicate that Mcm2-7 loading may be assisted by a natural predisposition of the hexamer to form open rings, and suggest a mechanism by which the CMG complex assists DNA strand separation. | |||||||||
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_1835.map.gz | 966.2 KB | EMDB map data format | |
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Header (meta data) | emd-1835-v30.xml emd-1835.xml | 11.7 KB 11.7 KB | Display Display | EMDB header |
Images | emd_1835.png | 120.9 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-1835 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-1835 | HTTPS FTP |
-Validation report
Summary document | emd_1835_validation.pdf.gz | 203.4 KB | Display | EMDB validaton report |
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Full document | emd_1835_full_validation.pdf.gz | 202.4 KB | Display | |
Data in XML | emd_1835_validation.xml.gz | 5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-1835 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-1835 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_1835.map.gz / Format: CCP4 / Size: 1001 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Lockwasher configuration of the Drosophila melanogaster Mcm2-7 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5.7 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 : Mcm2-7
Entire | Name: Mcm2-7 |
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Components |
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-Supramolecule #1000: Mcm2-7
Supramolecule | Name: Mcm2-7 / type: sample / ID: 1000 / Details: The sample was monodisperse / Oligomeric state: Hetero-hexamer / Number unique components: 6 |
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Molecular weight | Theoretical: 540 KDa |
-Macromolecule #1: Mcm2
Macromolecule | Name: Mcm2 / type: protein_or_peptide / ID: 1 / Name.synonym: Mcm2 / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Drosophila melanogaster (fruit fly) / synonym: Fruit Fly |
Molecular weight | Experimental: 100 KDa |
Recombinant expression | Organism: Baculovirus infected insect cells |
-Macromolecule #2: Mcm3
Macromolecule | Name: Mcm3 / type: protein_or_peptide / ID: 2 / Name.synonym: Mcm3 / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Drosophila melanogaster (fruit fly) / synonym: Fruit Fly |
Molecular weight | Experimental: 91 KDa |
Recombinant expression | Organism: Baculovirus infected insect cells |
-Macromolecule #3: Mcm4
Macromolecule | Name: Mcm4 / type: protein_or_peptide / ID: 3 / Name.synonym: Mcm4 / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Drosophila melanogaster (fruit fly) / synonym: Fruit Fly |
Molecular weight | Experimental: 97 KDa |
Recombinant expression | Organism: Baculovirus infected insect cells |
-Macromolecule #4: Mcm5
Macromolecule | Name: Mcm5 / type: protein_or_peptide / ID: 4 / Name.synonym: Mcm5 / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Drosophila melanogaster (fruit fly) / synonym: Fruit Fly |
Molecular weight | Experimental: 82 KDa |
Recombinant expression | Organism: Baculovirus infected insect cells |
-Macromolecule #5: Mcm6
Macromolecule | Name: Mcm6 / type: protein_or_peptide / ID: 5 / Name.synonym: Mcm6 / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Drosophila melanogaster (fruit fly) / synonym: Fruit Fly |
Molecular weight | Experimental: 92 KDa |
Recombinant expression | Organism: Baculovirus infected insect cells |
-Macromolecule #6: Mcm7
Macromolecule | Name: Mcm7 / type: protein_or_peptide / ID: 6 / Name.synonym: Mcm7 / Number of copies: 1 / Oligomeric state: Monomer / Recombinant expression: Yes |
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Source (natural) | Organism: Drosophila melanogaster (fruit fly) / synonym: Fruit Fly |
Molecular weight | Experimental: 81 KDa |
Recombinant expression | Organism: Baculovirus infected insect cells |
-Experimental details
-Structure determination
Processing | single particle reconstruction |
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Aggregation state | particle |
-Sample preparation
Vitrification | Cryogen name: NONE / Instrument: OTHER |
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-Electron microscopy
Microscope | FEI TECNAI 12 |
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Image recording | Average electron dose: 20 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: LAB6 |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal magnification: 30000 |
Sample stage | Specimen holder: Eucentric / Specimen holder model: SIDE ENTRY, EUCENTRIC |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 35.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: Spider |
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