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- EMDB-4788: D. melanogaster CMG-DNA with ATP, State 2B -

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Basic information

Entry
Database: EMDB / ID: EMD-4788
TitleD. melanogaster CMG-DNA with ATP, State 2B
Map dataState 2B, sharpened map (RELION PostProcess)
Sample
  • Complex: CMG helicase bound to forked DNA in the presence of ATP, State 2B
    • Complex: CMG helicase
      • Protein or peptide: x 11 types
    • Complex: DNA
      • DNA: x 2 types
  • Ligand: x 2 types
Function / homology
Function and homology information


Unwinding of DNA / Switching of origins to a post-replicative state / DNA endoreduplication / Assembly of the pre-replicative complex / Activation of ATR in response to replication stress / Activation of the pre-replicative complex / eggshell chorion gene amplification / Orc1 removal from chromatin / DNA amplification / DNA strand elongation involved in mitotic DNA replication ...Unwinding of DNA / Switching of origins to a post-replicative state / DNA endoreduplication / Assembly of the pre-replicative complex / Activation of ATR in response to replication stress / Activation of the pre-replicative complex / eggshell chorion gene amplification / Orc1 removal from chromatin / DNA amplification / DNA strand elongation involved in mitotic DNA replication / GINS complex / mitotic DNA replication preinitiation complex assembly / resolution of meiotic recombination intermediates / premeiotic DNA replication / mitotic DNA replication / CMG complex / MCM complex / DNA replication preinitiation complex / double-strand break repair via break-induced replication / mitotic DNA replication initiation / chromosome condensation / DNA strand elongation involved in DNA replication / DNA duplex unwinding / DNA unwinding involved in DNA replication / DNA replication origin binding / DNA replication initiation / DNA helicase activity / mitotic spindle organization / meiotic cell cycle / regulation of DNA-templated transcription elongation / helicase activity / single-stranded DNA binding / mitotic cell cycle / DNA helicase / DNA replication / cell division / chromatin binding / ATP hydrolysis activity / ATP binding / metal ion binding / nucleus / cytosol / cytoplasm
Similarity search - Function
CDC45 family / CDC45-like protein / DNA replication complex GINS protein Psf2 / GINS complex, subunit Psf1 / GINS complex, subunit Psf3 / GINS complex, subunit Psf3 superfamily / DNA replication complex GINS protein SLD5, C-terminal / GINS, helical bundle-like domain superfamily / GINS complex protein Sld5, alpha-helical domain / DNA replication complex GINS protein SLD5 C-terminus ...CDC45 family / CDC45-like protein / DNA replication complex GINS protein Psf2 / GINS complex, subunit Psf1 / GINS complex, subunit Psf3 / GINS complex, subunit Psf3 superfamily / DNA replication complex GINS protein SLD5, C-terminal / GINS, helical bundle-like domain superfamily / GINS complex protein Sld5, alpha-helical domain / DNA replication complex GINS protein SLD5 C-terminus / GINS complex subunit Sld5 / GINS subunit, domain A / GINS complex protein / MCM4, winged helix domain / DNA replication licensing factor Mcm5 / DNA replication licensing factor Mcm3 / Mini-chromosome maintenance complex protein 4 / DNA replication licensing factor Mcm6 / DNA replication licensing factor Mcm7 / Mcm6, C-terminal winged-helix domain / MCM6 C-terminal winged-helix domain / DNA replication licensing factor Mcm2 / Mini-chromosome maintenance protein 2 / Mini-chromosome maintenance, conserved site / MCM family signature. / MCM N-terminal domain / MCM N-terminal domain / MCM OB domain / MCM OB domain / Mini-chromosome maintenance protein / MCM, AAA-lid domain / MCM P-loop domain / MCM AAA-lid domain / MCM family domain profile. / minichromosome maintenance proteins / MCM domain / Winged helix-like DNA-binding domain superfamily / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / Nucleic acid-binding, OB-fold / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
CDC45L / DNA replication licensing factor Mcm2 / DNA replication licensing factor MCM4 / DNA replication licensing factor Mcm6 / DNA replication complex GINS protein SLD5 / DNA replication licensing factor Mcm5 / Probable DNA replication complex GINS protein PSF2 / DNA replication complex GINS protein PSF1 / DNA replication complex GINS protein PSF3 / DNA replication licensing factor Mcm7 / DNA replication licensing factor Mcm3
Similarity search - Component
Biological speciesDrosophila melanogaster (fruit fly)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.46 Å
AuthorsEickhoff P / Martino F / Locke J / Nans A / Costa A
CitationJournal: Cell Rep / Year: 2019
Title: Molecular Basis for ATP-Hydrolysis-Driven DNA Translocation by the CMG Helicase of the Eukaryotic Replisome.
Authors: Patrik Eickhoff / Hazal B Kose / Fabrizio Martino / Tatjana Petojevic / Ferdos Abid Ali / Julia Locke / Nele Tamberg / Andrea Nans / James M Berger / Michael R Botchan / Hasan Yardimci / Alessandro Costa /
Abstract: In the eukaryotic replisome, DNA unwinding by the Cdc45-MCM-Go-Ichi-Ni-San (GINS) (CMG) helicase requires a hexameric ring-shaped ATPase named minichromosome maintenance (MCM), which spools single- ...In the eukaryotic replisome, DNA unwinding by the Cdc45-MCM-Go-Ichi-Ni-San (GINS) (CMG) helicase requires a hexameric ring-shaped ATPase named minichromosome maintenance (MCM), which spools single-stranded DNA through its central channel. Not all six ATPase sites are required for unwinding; however, the helicase mechanism is unknown. We imaged ATP-hydrolysis-driven translocation of the CMG using cryo-electron microscopy (cryo-EM) and found that the six MCM subunits engage DNA using four neighboring protomers at a time, with ATP binding promoting DNA engagement. Morphing between different helicase states leads us to suggest a non-symmetric hand-over-hand rotary mechanism, explaining the asymmetric requirements of ATPase function around the MCM ring of the CMG. By imaging of a higher-order replisome assembly, we find that the Mrc1-Csm3-Tof1 fork-stabilization complex strengthens the interaction between parental duplex DNA and the CMG at the fork, which might support the coupling between DNA translocation and fork unwinding.
History
DepositionApr 8, 2019-
Header (metadata) releaseSep 18, 2019-
Map releaseSep 18, 2019-
UpdateJan 20, 2021-
Current statusJan 20, 2021Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0045
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.0045
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6raz
  • Surface level: 0.0045
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_4788.png

Downloads & links

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Map

FileDownload / File: emd_4788.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationState 2B, sharpened map (RELION PostProcess)
Voxel sizeX=Y=Z: 1.08 Å
Density
Contour LevelBy AUTHOR: 0.0045 / Movie #1: 0.0045
Minimum - Maximum-0.026608033 - 0.03887743
Average (Standard dev.)3.5806588e-05 (±0.00095414877)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 414.72003 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.081.081.08
M x/y/z384384384
origin x/y/z0.0000.0000.000
length x/y/z414.720414.720414.720
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS384384384
D min/max/mean-0.0270.0390.000

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Supplemental data

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Additional map: State 2B, non-sharpened map (RELION Refine3D)

Fileemd_4788_additional.map
AnnotationState 2B, non-sharpened map (RELION Refine3D)
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: State 2B, half-map 2 (RELION Refine3D)

Fileemd_4788_half_map_1.map
AnnotationState 2B, half-map 2 (RELION Refine3D)
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: State 2B, half-map 1 (RELION Refine3D)

Fileemd_4788_half_map_2.map
AnnotationState 2B, half-map 1 (RELION Refine3D)
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : CMG helicase bound to forked DNA in the presence of ATP, State 2B

EntireName: CMG helicase bound to forked DNA in the presence of ATP, State 2B
Components
  • Complex: CMG helicase bound to forked DNA in the presence of ATP, State 2B
    • Complex: CMG helicase
      • Protein or peptide: CDC45LCDC45-related protein
      • Protein or peptide: IP07275p
      • Protein or peptide: Probable DNA replication complex GINS protein PSF2
      • Protein or peptide: AT18545p
      • Protein or peptide: DNA replication complex GINS protein SLD5
      • Protein or peptide: DNA replication licensing factor Mcm2
      • Protein or peptide: DNA replication licensing factor Mcm5
      • Protein or peptide: DNA replication licensing factor Mcm6
      • Protein or peptide: DNA replication licensing factor Mcm3
      • Protein or peptide: DNA replication licensing factor MCM4
      • Protein or peptide: DNA replication licensing factor Mcm7
    • Complex: DNA
      • DNA: DNA
      • DNA: DNA
  • Ligand: ADENOSINE-5'-DIPHOSPHATE
  • Ligand: ADENOSINE-5'-TRIPHOSPHATE

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Supramolecule #1: CMG helicase bound to forked DNA in the presence of ATP, State 2B

SupramoleculeName: CMG helicase bound to forked DNA in the presence of ATP, State 2B
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#13

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Supramolecule #2: CMG helicase

SupramoleculeName: CMG helicase / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #3-#13
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)

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Supramolecule #3: DNA

SupramoleculeName: DNA / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #1-#2
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Recombinant expressionOrganism: synthetic construct (others)

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Macromolecule #1: DNA

MacromoleculeName: DNA / type: dna / ID: 1 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 6.389149 KDa
SequenceString:
(DC)(DG)(DT)(DT)(DT)(DT)(DA)(DT)(DT)(DT) (DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DA)(DA) (DA)

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Macromolecule #2: DNA

MacromoleculeName: DNA / type: dna / ID: 2 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 573.43 Da
SequenceString:
(DG)(DC)

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Macromolecule #3: CDC45L

MacromoleculeName: CDC45L / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 65.968789 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MFVQDLRNDF YRQLVGKRIL IVVNYDIDAI CASRILQALF KYDHMLYTVV PIMGVTGLKR AYGEHQGDVK YVVLVNCGGC VDIVELLQP SDDVTFFICD SHRPLDVCNI YSDRQVCILG DASLEENIPA FETIFYDSEG EDEDEDESSD TEQQHDDSGA G ESDQEDQA ...String:
MFVQDLRNDF YRQLVGKRIL IVVNYDIDAI CASRILQALF KYDHMLYTVV PIMGVTGLKR AYGEHQGDVK YVVLVNCGGC VDIVELLQP SDDVTFFICD SHRPLDVCNI YSDRQVCILG DASLEENIPA FETIFYDSEG EDEDEDESSD TEQQHDDSGA G ESDQEDQA PRSRKLSRLE RHEQRILKQR ARRQWESERD RIMFEYTQFS YYGRSAALMV FELAWKLSKD NMDLLWWAIV GI TEQLLLG KIESGAYTLE LEQIQSHVSR LTNKTNDQNT MSASKITFEN DLHLVLYRHW PVTESMRYSR YSSCQLKLWT LRG EKRLHE LLLEMGLPLV HARQTYGAMD LVLRKEFFSM VERLAEKYDI ADIVYGTFTL SYGYRSRYAA ADYVYALLAI MESV KKHKT PEDCFLEASD ALSRQHKQLL SAGIDQAKLL HAAVFRQVQS SLEARQVHSA GSFFYYVLQE EHAFFSYPYA LGLLA RFLL RGHVATSRAR QASDLPLIAS CPLNASEGMC LLVGIVPVRE DSPRNFFGKA FEQAAQKSGV ALLQDFFEPA VVQLRQ SDL TRFLDSLTVL LA

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Macromolecule #4: IP07275p

MacromoleculeName: IP07275p / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 23.333693 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSRQTKMFGE KAFDLLKELE RSSQTIPAFD DDGVRQVLEE IKAIFEENVA QASSYNASGD RSLWPLLNFR HAALQRNKRC LLAYLYERC RRIKALRWEF GPIIPGDIKQ ALCEPEVTFF NNYSKSLAAY MCSAGYNQGL PIDLTNNLRP PKSLYIEVRC M EDYGKFEL ...String:
MSRQTKMFGE KAFDLLKELE RSSQTIPAFD DDGVRQVLEE IKAIFEENVA QASSYNASGD RSLWPLLNFR HAALQRNKRC LLAYLYERC RRIKALRWEF GPIIPGDIKQ ALCEPEVTFF NNYSKSLAAY MCSAGYNQGL PIDLTNNLRP PKSLYIEVRC M EDYGKFEL DDGEVIHLKK NSQHYLPRAQ VESLVRQGIL HHIA

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Macromolecule #5: Probable DNA replication complex GINS protein PSF2

MacromoleculeName: Probable DNA replication complex GINS protein PSF2 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 23.141781 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDPSIIEFIG EKCMISIIPN FSNEPLHLIY GPVGPFRAGF PVFVPLWMAT HLRKQQKCRI VPPEWMDMDI LEEIKEEEKR SKFFTKMPC EHYMVVAQLV MSTAPDDVPR CEELRTVIKD IFDIRESKLR TSIDAFIKGE GTYAKLDNLT LLEIHSVRPI L PYSLDHIA ...String:
MDPSIIEFIG EKCMISIIPN FSNEPLHLIY GPVGPFRAGF PVFVPLWMAT HLRKQQKCRI VPPEWMDMDI LEEIKEEEKR SKFFTKMPC EHYMVVAQLV MSTAPDDVPR CEELRTVIKD IFDIRESKLR TSIDAFIKGE GTYAKLDNLT LLEIHSVRPI L PYSLDHIA RYQRTATASQ RDTSMLSASM AGSSSGPNSN SLFSQ

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Macromolecule #6: AT18545p

MacromoleculeName: AT18545p / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 24.829984 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MNGMNYFPNY YSIEDIFVTQ EKVECRVNTK LQRMGFLDSG AESDDLEPGR TVNLPLWYIK ELKVNNAYFT VAVPDIYRNV HKAVCEAET THIELGRLHP YFYEFGRYLT PYDRNHVIGR IIFETLRQRV RHLLDISKSD GQAAKAEHRL DNIEAKLHEA G VRTNSQYI ...String:
MNGMNYFPNY YSIEDIFVTQ EKVECRVNTK LQRMGFLDSG AESDDLEPGR TVNLPLWYIK ELKVNNAYFT VAVPDIYRNV HKAVCEAET THIELGRLHP YFYEFGRYLT PYDRNHVIGR IIFETLRQRV RHLLDISKSD GQAAKAEHRL DNIEAKLHEA G VRTNSQYI EWLQMTGNKI RTSELVEEHQ KKRRRADRSD DEGDALPNSK RATL

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Macromolecule #7: DNA replication complex GINS protein SLD5

MacromoleculeName: DNA replication complex GINS protein SLD5 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 26.148234 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSDVEDVPET QLEIDVSDGA GLEDEDDDDM EQITAQKVLE IIETAWINEM CAPEILPSQT DMLELMVSQV AHMEEQMRDL DKNDFRAVV HSMELERVRY IMASYLRCRL QKIETFTQHI LNQEESREPD DKRLSPEETK FAQEFASNVD EYFHKVATQY M PNQQRGEA ...String:
MSDVEDVPET QLEIDVSDGA GLEDEDDDDM EQITAQKVLE IIETAWINEM CAPEILPSQT DMLELMVSQV AHMEEQMRDL DKNDFRAVV HSMELERVRY IMASYLRCRL QKIETFTQHI LNQEESREPD DKRLSPEETK FAQEFASNVD EYFHKVATQY M PNQQRGEA EQRIVTPNLM SHVFLKANVA VPAVIVGVDD EEVDMAAGSQ HIIPYQLVAD LIQNNQAQLI

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Macromolecule #8: DNA replication licensing factor Mcm2

MacromoleculeName: DNA replication licensing factor Mcm2 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 100.537438 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDNPSSPPPN TPSDAAERRD LRAAMTSPVG DFEPFENEDE ILGDQTVRDE AEEEDGEELF GDNMENDYRP MPELDHYDPA LLDDEDDFS EMSQGDRFAA ESEMRRRDRA AGIHRDDRDL GFGQSDDEDD VGPRAKRRAG EKAAVGEVED TEMVESIENL E DTKGHSTK ...String:
MDNPSSPPPN TPSDAAERRD LRAAMTSPVG DFEPFENEDE ILGDQTVRDE AEEEDGEELF GDNMENDYRP MPELDHYDPA LLDDEDDFS EMSQGDRFAA ESEMRRRDRA AGIHRDDRDL GFGQSDDEDD VGPRAKRRAG EKAAVGEVED TEMVESIENL E DTKGHSTK EWVSMLGPRT EIANRFQSFL RTFVDERGAY TYRDRIRRMC EQNMSSFVVS YTDLANKEHV LAYFLPEAPF QM LEIFDKV AKDMVLSIFP TYERVTTEIH VRISELPLIE ELRTFRKLHL NQLVRTLGVV TATTGVLPQL SVIKYDCVKC GYV LGPFVQ SQNTEIKPGS CPECQSTGPF SINMEQTLYR NYQKITLQES PGRIPAGRIP RSKDVILLAD LCDQCKPGDE LEVT GIYTN NYDGSLNTDQ GFPVFATVII ANHVVVKDSK QVVQSLTDED IATIQKLSKD PRIVERVVAS MAPSIYGHDY IKRAL ALAL FGGESKNPGE KHKVRGDINL LICGDPGTAK SQFLKYTEKV APRAVFTTGQ GASAVGLTAY VRRNPVSREW TLEAGA LVL ADQGVCLIDE FDKMNDQDRT SIHEAMEQQS ISISKAGIVT SLQARCTVIA AANPIGGRYD PSMTFSENVN LSEPILS RF DVLCVVKDEF DPMQDQQLAK FVVHSHMKHH PSEEEQPELE EPQLKTVDEI PQDLLRQYIV YAKENIRPKL TNIDEDKI A KMYAQLRQES FATGSLPITV RHIESVIRMS EAHARMHLRE NVMEADVSMA IRMMLESFIE AQKFSVMKKM RSTFQKYLS FQKDHSELLF FILRQLTLDQ LAYIRCKDGP GATHVEIMER DLIERAKQLD IVNLKPFYES DLFRTNGFSY DPKRRIILQI VVDGNTA

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Macromolecule #9: DNA replication licensing factor Mcm5

MacromoleculeName: DNA replication licensing factor Mcm5 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 82.37532 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MEGFDDAGVF FSDNFGGDNQ QDAQINLQAV KKKYKEFIRT FNEENFFYKY RDTLKRNYLN GRYFLEIEME DLVGFDETLA DKLNKQPTE HLEIFEEAAR EVADEITAPR PEHEEHMHDI QILLSSNANP TNIRQLKSDC VSKLVKIAGI IVAASGISAK A TRMSIQCL ...String:
MEGFDDAGVF FSDNFGGDNQ QDAQINLQAV KKKYKEFIRT FNEENFFYKY RDTLKRNYLN GRYFLEIEME DLVGFDETLA DKLNKQPTE HLEIFEEAAR EVADEITAPR PEHEEHMHDI QILLSSNANP TNIRQLKSDC VSKLVKIAGI IVAASGISAK A TRMSIQCL SCSTVIPNLK VNPGLEGYAL PRKCNTEQAG RPKCPLDPFF IMPDKCKCVD FQTLKLQELP DFVPQGEIPR HL QLFCDRS LCERVVPGNR VLIQGIYSIR KVGKPSRRDG REKAVVGVRA PYMRVVGITV DSEGAGAISR YSNITSDEEE HFR RMAASG DIYERLSQSL APSIFGSRDI KKAITCMLFG GSRKRLPDGL CRRGDINVLL LGDPGTAKSQ LLKFVEKVAP IAVY TSGKG SSAAGLTASV MKDPQTRNFV MEGGAMVLAD GGVVCIDEFD KMREDDRVAI HEAMEQQTIS IAKAGITTTL NSRCS VLAA ANSIFGRWDD TKGEENIDFM PTILSRFDMI FIVKDIHDES RDITLAKHII NVHLSSNKSA PSEPAEGEIS LSTFKK YIH YCRTHCGPRL SEAAGEKLKS RYVLMRSGAG QQEKASDKRL SIPITVRQLE AVIRISESLA KIRLQPFATD EHVNEAL RL FQVSTLDAAM TGSLAGAEGF TTEEDQETLN RIEKQLKRRF AIGSQVSEQN ILQDFLRQKY EERTVMKVIH TMIRRGEL Q HRMQRKMLYR IC

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Macromolecule #10: DNA replication licensing factor Mcm6

MacromoleculeName: DNA replication licensing factor Mcm6 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 92.46782 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDVADAQVGQ LRVKDEVGIR AQKLFQDFLE EFKEDGEIKY TRPAASLESP DRCTLEVSFE DVEKYDQNLA TAIIEEYYHI YPFLCQSVS NYVKDRIGLK TQKDCYVAFT EVPTRHKVRD LTTSKIGTLI RISGQVVRTH PVHPELVSGV FMCLDCQTEI R NVEQQFKF ...String:
MDVADAQVGQ LRVKDEVGIR AQKLFQDFLE EFKEDGEIKY TRPAASLESP DRCTLEVSFE DVEKYDQNLA TAIIEEYYHI YPFLCQSVS NYVKDRIGLK TQKDCYVAFT EVPTRHKVRD LTTSKIGTLI RISGQVVRTH PVHPELVSGV FMCLDCQTEI R NVEQQFKF TNPTICRNPV CSNRKRFMLD VEKSLFLDFQ KIRIQETQAE LPRGCIPRAV EIILRSELVE TVQAGDRYDF TG TLIVVPD VSVLAGVGTR AENSSRHKPG EGMDGVTGLK ALGMRELNYR MAFLACSVQA TTARFGGTDL PMSEVTAEDM KKQ MTDAEW HKIYEMSKDR NLYQNLISSL FPSIYGNDEV KRGILLQQFG GVAKTTTEKT SLRGDINVCI VGDPSTAKSQ FLKQ VSDFS PRAIYTSGKA SSAAGLTAAV VRDEESFDFV IEAGALMLAD NGICCIDEFD KMDQRDQVAI HEAMEQQTIS IARAG VRAT LNARTSILAA ANPINGRYDR SKSLQQNIQL SAPIMSRFDL FFILVDECNE VVDYAIARKI VDLHSNIEES VERAYT REE VLRYVTFARQ FKPVISQEAG HMLVENYGHL RQRDTGTSGR STWRITVRQL ESMIRLSEAM AKLECSNRVL ERHVKEA FR LLNKSIIRVE QPDIHLDDDE GLDMDDGIQH DIDMENNGAA ANVDENNGMD TSASGAVQKK KFTLSFEDYK NLSTMLVL H MRAEEARCEV EGNDTGIKRS NVVTWYLEQV ADQIESEDEL ISRKNLIEKL IDRLIYHDQV IIPLKTSTLK PRIQVQKDF VEEDDPLLVV HPNYVVE

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Macromolecule #11: DNA replication licensing factor Mcm3

MacromoleculeName: DNA replication licensing factor Mcm3 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 91.045164 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MAHEGEQFIK DIQREYVDFL DDEEDQGIYA GHVKDMIAEK SKRLIVNVND LKRKNPQRAL GLLSNAADEQ LAFGRALKEY ASTVDPGYA KMHEDLFVGF EGCFGNRHVT PRSLTSIYLG NMVCVEGIVT KVSLIRPKVV RSVHYCPNTR KVMERKYTDL T SFEAVPSG ...String:
MAHEGEQFIK DIQREYVDFL DDEEDQGIYA GHVKDMIAEK SKRLIVNVND LKRKNPQRAL GLLSNAADEQ LAFGRALKEY ASTVDPGYA KMHEDLFVGF EGCFGNRHVT PRSLTSIYLG NMVCVEGIVT KVSLIRPKVV RSVHYCPNTR KVMERKYTDL T SFEAVPSG AAYPTKDEDG NLLETEYGLS VYKDHQTLTI QEMPEKAPAG QLPRSVDIVC DDDLVDRCKP GDRVQIVGSY RC LPGKRGG YTSGTFRTVL LANNISLLSK ESNLDISRED IMLCKKLAKN NDIFELLSKS LAPSIHGHAY VKQAILCLLL GGV EKILPN GTRLRGDINV LLIGDPSVAK SQLLRYVLNT APRAIPTTGR GSSGVGLTAA VTTDQETGER RLEAGAMVLA DRGV VCIDE FDKMSDIDRT AIHEVMEQGR VTISKAGIHA SLNARCSVLA AANPVYGRYD QYKTPMENIG LQDSLLSRFD LLFVM LDVI DSDVDQMISD HVVRMHRYRN PKEADGEPLS MGSSYADSLS FVSSSEEKKD TEVYEKYDAL LHGKSRQRHE KILSVE FMR KYIHIAKCMK PKLGEQACEA IANEYSRLRS QEAVETDVAR TQPITARTLE TLIRLSTAHA RARMSKSVTI DDAHAAI EL VQFAYFKKVL DKDRPSKRRR NSGSDAEDDN GEASSQRSPS RRSKRTRTAT VGADSDEEDI EPPQPDAGDL TRRETRRS L PARSVAMLMA SPSSEEQSVA TSTTEPAIIS DARLGEFKNN LQRLFREARE QSLALARITT AINVGSQEPF TAGEIEAAV HRMTEDNQIM VADDIVFLI

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Macromolecule #12: DNA replication licensing factor MCM4

MacromoleculeName: DNA replication licensing factor MCM4 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 96.735094 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSSPARSPSV GGATPKQGAR TPTRGIASQD VETPMRMGPG RAVRPSDNIS LPPTSPGNIS LPATSPARGL GANMSEIDLS SPLNYGTPS SMGSIRTPRS GIRGTPLRAR PDIRTDKRIR QVAIGGGSGL EPIPEKGSET TDPVSESSQA PQLVVWGTNV V VSQCKSKF ...String:
MSSPARSPSV GGATPKQGAR TPTRGIASQD VETPMRMGPG RAVRPSDNIS LPPTSPGNIS LPATSPARGL GANMSEIDLS SPLNYGTPS SMGSIRTPRS GIRGTPLRAR PDIRTDKRIR QVAIGGGSGL EPIPEKGSET TDPVSESSQA PQLVVWGTNV V VSQCKSKF KSFIMRFIDP SAEQDEISEN IDVNQPLYLQ KLEEIHTLEE PYLNLNCAHL KTFDQDLYRQ LICYPQEVIP GF DMAINEM FFERYPAALL EHQIQVRPFN ADKTRNMRSL NPEDMDQLIS ISGMVIRSSN VIPEMREAFF SCNICSFSTT VEV DRGRIN QPTLCTNCNT NHCFRLIHNR SEFTDKQLVK LQESPDDMAA GQTPHNVLLY AHNDLVDKVQ PGDRVTVTGI YRAT PLKTG GLSSSVKSVY KTHVDVVHFR KVDNKRLYED EEGKDHIFPP ERVELLQLLA KKPDIYDRLA RAIAPSIYEN DDIKK GILL QLFGGTKKKH ATLGRQNFRS EIHLLLCGDP GTSKSQMLQY VFNLVPRSQY TSGRGSSAVG LTAYVTKDPE TRQLVL QTG ALVLADNGVC CIDEFDKMND STRSVLHEVM EQQTLSIAKA GIICQLNART SILAAANPAE SQWNKRKNII DNVQLPH TL LSRFDLIFLV LDPQDEIFDK RLASHLVSLY YVTRHEEEDT MFDMSVLRDY IAYAREHLSP TLSDEAQQRL IQAYVDMR K VGAGRGQISA YPRQLESLIR LSEAHAKVRL SNQVELLDVE EAWRLHREAL KQSATDPLSG KIDVGILTTG LSTAARKKR ADLVAAIKEN LKKKGKVLTV PYQKLFSDIK EGSQIMITRE QFEDALKEVQ DEGAIVVMGK NTIRIC

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Macromolecule #13: DNA replication licensing factor Mcm7

MacromoleculeName: DNA replication licensing factor Mcm7 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Drosophila melanogaster (fruit fly)
Molecular weightTheoretical: 81.399352 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MARRDYAQDR ESIKTFLSEF CKCDDDGKKE FVYGSQLVKL AHREQVLITI DLDDLAEFNE SLAEAVVDNC RRYTSIFSDV IAELLPSYK QQEVHAKDAL DVYIEHRLMM ESRTRNPMEQ RDERNSFPSE LMKRFEVGFK PLSTEKAHSI REVKAQHIGK L VTVRGIVT ...String:
MARRDYAQDR ESIKTFLSEF CKCDDDGKKE FVYGSQLVKL AHREQVLITI DLDDLAEFNE SLAEAVVDNC RRYTSIFSDV IAELLPSYK QQEVHAKDAL DVYIEHRLMM ESRTRNPMEQ RDERNSFPSE LMKRFEVGFK PLSTEKAHSI REVKAQHIGK L VTVRGIVT RCTEVKPMMV VATYTCDRCG SETYQPVNSL SFTPVHDCPS DDCRVNKAGG RLYLQTRGSK FVKFQEVKMQ EH SDQVPVG HIPRSMTIMC RGEVTRMAQP GDHIVVSGVF LPLMRTGFAQ MIQGLLSETF LQAHRIICIN KNDEISDKDA ELT PEELEE LAQDDFYERL ATSLAPEIYG HLDVKKALLL LLVGGVDKRP DGMKIRGNIN ICLMGDPGVA KSQLLGYISR LAVR SQYTT GRGSSGVGLT AAVMKDPLTG EMTLEGGALV LADQGVCCID EFDKMADQDR TAIHEVMEQQ TISIAKAGIM TTLNA RVSI LAAANPAFGR YNPRRTVEQN IQLPAALLSR FDLLWLIQDK PDRDNDLRLA KHITYVHSHS KQPPTRVKAL DMNLMR RYI NLCKRKNPTI PDELTDYIVG AYVELRREAR NQKDMTFTSA RNLLGILRLS TALARLRLSD SVEKDDVAEA LRLLEMS KD SLNQIHEHQK GHVPNTSDRI FAIVRELAGS GKAVKISDIM DRCTTKGFKP DQVDKCIDDY EELNVWQVNM GRTKITFM

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Macromolecule #14: ADENOSINE-5'-DIPHOSPHATE

MacromoleculeName: ADENOSINE-5'-DIPHOSPHATE / type: ligand / ID: 14 / Number of copies: 2 / Formula: ADP
Molecular weightTheoretical: 427.201 Da
Chemical component information

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

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Macromolecule #15: ADENOSINE-5'-TRIPHOSPHATE

MacromoleculeName: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 15 / Number of copies: 4 / Formula: ATP
Molecular weightTheoretical: 507.181 Da
Chemical component information

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM / Adenosine triphosphate

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.6
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.46 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 61082
FSC plot (resolution estimation)

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