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- EMDB-15309: Pol alpha - replisome complex containing Ctf4. Engaged on a DNA f... -
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Open data
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Basic information
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Title | Pol alpha - replisome complex containing Ctf4. Engaged on a DNA fork containing a 60 nucleotide lagging strand. | |||||||||
![]() | Pol alpha - replisome complex containing Ctf4. Unsharpened. | |||||||||
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![]() | Replication / helicase / polymerase / pol alpha / priming | |||||||||
Function / homology | ![]() Inhibition of replication initiation of damaged DNA by RB1/E2F1 / establishment of sister chromatid cohesion / maintenance of DNA repeat elements / Unwinding of DNA / replication fork arrest / regulation of nuclear cell cycle DNA replication / Cul8-RING ubiquitin ligase complex / RNA-templated DNA biosynthetic process / DNA replication initiation / meiotic chromosome segregation ...Inhibition of replication initiation of damaged DNA by RB1/E2F1 / establishment of sister chromatid cohesion / maintenance of DNA repeat elements / Unwinding of DNA / replication fork arrest / regulation of nuclear cell cycle DNA replication / Cul8-RING ubiquitin ligase complex / RNA-templated DNA biosynthetic process / DNA replication initiation / meiotic chromosome segregation / Processive synthesis on the lagging strand / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / Removal of the Flap Intermediate / GINS complex / DNA strand elongation involved in mitotic DNA replication / nuclear DNA replication / mitotic DNA replication preinitiation complex assembly / Polymerase switching / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / alpha DNA polymerase:primase complex / mitotic DNA replication / anaphase-promoting complex binding / Activation of the pre-replicative complex / CMG complex / establishment of mitotic sister chromatid cohesion / DNA replication checkpoint signaling / single-stranded 3'-5' DNA helicase activity / entrainment of circadian clock / nuclear pre-replicative complex / DNA primase activity / MCM complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / lagging strand elongation / DNA replication, synthesis of primer / replication fork protection complex / telomere capping / mitotic DNA replication checkpoint signaling / mitotic DNA replication initiation / double-strand break repair via break-induced replication / mitotic intra-S DNA damage checkpoint signaling / silent mating-type cassette heterochromatin formation / regulation of DNA-templated DNA replication initiation / single-stranded DNA helicase activity / mitotic sister chromatid cohesion / DNA strand elongation involved in DNA replication / nuclear chromosome / DNA synthesis involved in DNA repair / DNA biosynthetic process / leading strand elongation / replication fork processing / DNA unwinding involved in DNA replication / nuclear replication fork / mitotic G2 DNA damage checkpoint signaling / 3'-5' DNA helicase activity / DNA replication origin binding / subtelomeric heterochromatin formation / DNA replication initiation / Ub-specific processing proteases / heterochromatin formation / telomere maintenance / nuclear periphery / helicase activity / meiotic cell cycle / replication fork / DNA-templated DNA replication / double-strand break repair / Transferases; Transferring phosphorus-containing groups; Nucleotidyltransferases / nucleosome assembly / nuclear envelope / mitotic cell cycle / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / DNA replication / DNA helicase / chromosome, telomeric region / DNA-directed DNA polymerase / DNA-directed DNA polymerase activity / DNA repair / nucleotide binding / DNA damage response / chromatin binding / ATP hydrolysis activity / mitochondrion / DNA binding / nucleoplasm / ATP binding / identical protein binding / nucleus / metal ion binding / cytoplasm / cytosol Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.64 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 | ![]() | 20.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 13.9 KB 13.9 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 7.2 KB | Display | ![]() |
Images | ![]() | 100.9 KB | ||
Others | ![]() ![]() ![]() | 2 MB 37.7 MB 37.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1 MB | Display | ![]() |
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Full document | ![]() | 1 MB | Display | |
Data in XML | ![]() | 14.2 KB | Display | |
Data in CIF | ![]() | 18.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8b9aMC ![]() 8b9bMC ![]() 8b9cC ![]() 8b9dC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Pol alpha - replisome complex containing Ctf4. Unsharpened. | ||||||||||||||||||||
Voxel size | X=Y=Z: 2.26727 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Locally filtered map
File | emd_15309_additional_1.map | ||||||||||||
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Annotation | Locally filtered map | ||||||||||||
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Density Histograms |
-Half map: Half map
File | emd_15309_half_map_1.map | ||||||||||||
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Annotation | Half map | ||||||||||||
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Density Histograms |
-Half map: Half map
File | emd_15309_half_map_2.map | ||||||||||||
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Annotation | Half map | ||||||||||||
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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 |