- EMDB-19187: Cryo-EM structure of an MCM helicase single hexamer loaded onto dsDNA. -
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Open data
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Basic information
Entry
Database: EMDB / ID: EMD-19187
Title
Cryo-EM structure of an MCM helicase single hexamer loaded onto dsDNA.
Map data
Primary map from final 3D refinement in Relion.
Sample
Complex: Hexameric MCM helicase complex loaded onto duplex DNA
Protein or peptide: x 6 types
DNA: x 2 types
Ligand: x 4 types
Keywords
MCM helicase / DNA replication / REPLICATION
Function / homology
Function and homology information
MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / mitotic DNA replication / Activation of the pre-replicative complex / CMG complex ...MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / mitotic DNA replication / Activation of the pre-replicative complex / CMG complex / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / MCM complex / replication fork protection complex / double-strand break repair via break-induced replication / single-stranded DNA helicase activity / mitotic DNA replication initiation / silent mating-type cassette heterochromatin formation / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / nuclear replication fork / DNA replication origin binding / DNA replication initiation / subtelomeric heterochromatin formation / helicase activity / transcription elongation by RNA polymerase II / heterochromatin formation / single-stranded DNA binding / DNA helicase / forked DNA-dependent helicase activity / single-stranded 3'-5' DNA helicase activity / four-way junction helicase activity / double-stranded DNA helicase activity / chromosome, telomeric region / DNA replication / DNA damage response / chromatin binding / ATP hydrolysis activity / zinc ion binding / nucleoplasm / ATP binding / nucleus / cytoplasm Similarity search - Function
Macromolecule #2: DNA replication licensing factor MCM3
Macromolecule
Name: DNA replication licensing factor MCM3 / type: protein_or_peptide / ID: 2 Details: MCM3 contains a CBP-TEV tag at the N-terminus: KRRWKKNFIAVSAANRFKKISSSGALENLYFQG Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GRAPHENE OXIDE / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 300 sec. / Pretreatment - Atmosphere: AIR Details: Glow discharge at 40 mA for 5 minutes prior to applying graphene oxide.
Vitrification
Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 283 K / Instrument: FEI VITROBOT MARK IV
Details
Complete biochemical reaction containing the MCM helicase in complex with Cdt1, the helicase loader (the origin recognition complex), the co-factor Cdc6, an origin or replication (DNA) flanked by nucleosomes, the regulatory kinase CDK and an inhibitor of CDK (Sic1). The sample was incubated with ATP yielding a range of structural intermediates in the MCM loading pathway.
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Electron microscopy
Microscope
FEI TITAN KRIOS
Specialist optics
Energy filter - Slit width: 20 eV
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-40 / Number real images: 24506 / Average electron dose: 51.2 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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