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- EMDB-15340: MCM C-tier local refinement. Human replisome bound by pol alpha, ... -
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Open data
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Basic information
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Title | MCM C-tier local refinement. Human replisome bound by pol alpha, engaged on fork DNA with 60 nt lagging strand. | |||||||||
![]() | MCM C-tier local refinement. Human replisome - pol alpha complex. Engaged with fork DNA substrate with 60 nt lagging strand. Sharp | |||||||||
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![]() | Replication / helicase / polymerase / pol alpha / priming | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | |||||||||
![]() | Jones ML / Yeeles JTP | |||||||||
Funding support | ![]()
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![]() | ![]() Title: How Pol α-primase is targeted to replisomes to prime eukaryotic DNA replication. Authors: Morgan L Jones / Valentina Aria / Yasemin Baris / Joseph T P Yeeles / ![]() Abstract: During eukaryotic DNA replication, Pol α-primase generates primers at replication origins to start leading-strand synthesis and every few hundred nucleotides during discontinuous lagging-strand ...During eukaryotic DNA replication, Pol α-primase generates primers at replication origins to start leading-strand synthesis and every few hundred nucleotides during discontinuous lagging-strand replication. How Pol α-primase is targeted to replication forks to prime DNA synthesis is not fully understood. Here, by determining cryoelectron microscopy (cryo-EM) structures of budding yeast and human replisomes containing Pol α-primase, we reveal a conserved mechanism for the coordination of priming by the replisome. Pol α-primase binds directly to the leading edge of the CMG (CDC45-MCM-GINS) replicative helicase via a complex interaction network. The non-catalytic PRIM2/Pri2 subunit forms two interfaces with CMG that are critical for in vitro DNA replication and yeast cell growth. These interactions position the primase catalytic subunit PRIM1/Pri1 directly above the exit channel for lagging-strand template single-stranded DNA (ssDNA), revealing why priming occurs efficiently only on the lagging-strand template and elucidating a mechanism for Pol α-primase to overcome competition from RPA to initiate primer synthesis. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 116.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 16.2 KB 16.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 10.5 KB | Display | ![]() |
Images | ![]() | 148.4 KB | ||
Others | ![]() ![]() ![]() ![]() | 118 MB 60.4 MB 116.1 MB 116.1 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 965 KB | Display | ![]() |
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Full document | ![]() | 964.5 KB | Display | |
Data in XML | ![]() | 18.6 KB | Display | |
Data in CIF | ![]() | 24 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | MCM C-tier local refinement. Human replisome - pol alpha complex. Engaged with fork DNA substrate with 60 nt lagging strand. Sharp | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.2363 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: MCM C-tier local refinement. Human replisome - pol...
File | emd_15340_additional_1.map | ||||||||||||
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Annotation | MCM C-tier local refinement. Human replisome - pol alpha complex. Engaged with fork DNA substrate with 60 nt lagging strand. Sharp | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: MCM C-tier local refinement. Human replisome - pol...
File | emd_15340_additional_2.map | ||||||||||||
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Annotation | MCM C-tier local refinement. Human replisome - pol alpha complex. Engaged with fork DNA substrate with 60 nt lagging strand. Unsharp | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map B
File | emd_15340_half_map_1.map | ||||||||||||
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Annotation | Half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map A
File | emd_15340_half_map_2.map | ||||||||||||
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Annotation | Half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Replisome - pol alpha complex
Entire | Name: Replisome - pol alpha complex |
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Components |
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-Supramolecule #1: Replisome - pol alpha complex
Supramolecule | Name: Replisome - pol alpha complex / type: complex / ID: 1 / Parent: 0 Details: S. cerevisiae pol alpha bound to the core replisome engaged with a fork DNA substrate containing a 60 nucleotide lagging strand. |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.6 |
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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: 40.184 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |