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- EMDB-18195: Single CMG purified from replicating Xenopus egg extracts -

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Basic information

Entry
Database: EMDB / ID: EMD-18195
TitleSingle CMG purified from replicating Xenopus egg extracts
Map data
Sample
  • Complex: Single CMG purified from replicating Xenopus egg extracts
    • Protein or peptide: x 11 types
KeywordsDNA replication initiation / Xenopus egg extract / primordial dwarfism / replicative helicase / genome stability / REPLICATION
Function / homology
Function and homology information


GINS complex / premeiotic DNA replication / CMG complex / MCM complex / regulation of DNA-templated DNA replication initiation / DNA unwinding involved in DNA replication / DNA replication origin binding / DNA replication initiation / DNA helicase activity / helicase activity ...GINS complex / premeiotic DNA replication / CMG complex / MCM complex / regulation of DNA-templated DNA replication initiation / DNA unwinding involved in DNA replication / DNA replication origin binding / DNA replication initiation / DNA helicase activity / helicase activity / DNA-templated DNA replication / DNA helicase / DNA replication / cell cycle / chromatin binding / chromatin / ATP hydrolysis activity / DNA binding / nucleoplasm / ATP binding / metal ion binding / nucleus / cytoplasm
Similarity search - Function
CDC45 family / CDC45-like protein / 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 ...CDC45 family / CDC45-like protein / 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 / 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 / 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
DNA replication licensing factor mcm4-B / Maternal DNA replication licensing factor mcm3 / DNA replication licensing factor mcm2 / DNA replication licensing factor mcm5-A / Maternal DNA replication licensing factor mcm6 / DNA replication complex GINS protein PSF3 / DNA replication complex GINS protein PSF1 / DNA replication complex GINS protein SLD5 / DNA replication licensing factor mcm7-B / Cell division control protein 45 homolog
Similarity search - Component
Biological speciesXenopus laevis (African clawed frog)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.03 Å
AuthorsCvetkovic MA / Costa A
Funding support United Kingdom, European Union, 5 items
OrganizationGrant numberCountry
The Francis Crick InstituteFC001065 United Kingdom
European Research Council (ERC)820102European Union
Marie Sklodowska-Curie Actions, FragNET ITN845939European Union
Wellcome Trust215510/Z/19/Z United Kingdom
Biotechnology and Biological Sciences Research Council (BBSRC)BB/T001860/1 United Kingdom
CitationJournal: Mol Cell / Year: 2023
Title: The structural mechanism of dimeric DONSON in replicative helicase activation.
Authors: Milos A Cvetkovic / Paolo Passaretti / Agata Butryn / Alicja Reynolds-Winczura / Georgia Kingsley / Aggeliki Skagia / Cyntia Fernandez-Cuesta / Divyasree Poovathumkadavil / Roger George / ...Authors: Milos A Cvetkovic / Paolo Passaretti / Agata Butryn / Alicja Reynolds-Winczura / Georgia Kingsley / Aggeliki Skagia / Cyntia Fernandez-Cuesta / Divyasree Poovathumkadavil / Roger George / Anoop S Chauhan / Satpal S Jhujh / Grant S Stewart / Agnieszka Gambus / Alessandro Costa /
Abstract: The MCM motor of the replicative helicase is loaded onto origin DNA as an inactive double hexamer before replication initiation. Recruitment of activators GINS and Cdc45 upon S-phase transition ...The MCM motor of the replicative helicase is loaded onto origin DNA as an inactive double hexamer before replication initiation. Recruitment of activators GINS and Cdc45 upon S-phase transition promotes the assembly of two active CMG helicases. Although work with yeast established the mechanism for origin activation, how CMG is formed in higher eukaryotes is poorly understood. Metazoan Downstream neighbor of Son (DONSON) has recently been shown to deliver GINS to MCM during CMG assembly. What impact this has on the MCM double hexamer is unknown. Here, we used cryoelectron microscopy (cryo-EM) on proteins isolated from replicating Xenopus egg extracts to identify a double CMG complex bridged by a DONSON dimer. We find that tethering elements mediating complex formation are essential for replication. DONSON reconfigures the MCM motors in the double CMG, and primordial dwarfism patients' mutations disrupting DONSON dimerization affect GINS and MCM engagement in human cells and DNA synthesis in Xenopus egg extracts.
History
DepositionAug 14, 2023-
Header (metadata) releaseOct 18, 2023-
Map releaseOct 18, 2023-
UpdateNov 29, 2023-
Current statusNov 29, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_18195.png

Downloads & links

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Map

FileDownload / File: emd_18195.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 0.95 Å
Density
Contour LevelBy AUTHOR: 0.4
Minimum - Maximum-1.8589205 - 3.0845175
Average (Standard dev.)0.00003052095 (±0.06454463)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions480480480
Spacing480480480
CellA=B=C: 456.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_18195_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_18195_half_map_2.map
Projections & Slices
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Slices (1/2)
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Histogram

Histogram (log scale)

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Sample components

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Entire : Single CMG purified from replicating Xenopus egg extracts

EntireName: Single CMG purified from replicating Xenopus egg extracts
Components
  • Complex: Single CMG purified from replicating Xenopus egg extracts
    • Protein or peptide: DNA replication licensing factor Mcm2 - Xenopus laevis
    • Protein or peptide: DNA replication licensing factor Mcm3 - Xenopus laevis
    • Protein or peptide: DNA replication licensing factor Mcm4 - Xenopus laevis
    • Protein or peptide: DNA replication licensing factor Mcm5 - Xenopus laevis
    • Protein or peptide: DNA replication licensing factor Mcm6 - Xenopus laevis
    • Protein or peptide: DNA replication licensing factor Mcm7 - Xenopus laevis
    • Protein or peptide: DNA replication complex GINS protein Psf1 - Xenopus laevis
    • Protein or peptide: DNA replication complex GINS protein Psf2 - Xenopus laevis
    • Protein or peptide: DNA replication complex GINS protein Psf3 - Xenopus laevis
    • Protein or peptide: DNA replication complex GINS protein Sld5 - Xenopus laevis
    • Protein or peptide: Cell division control protein 45 - Xenopus laevis

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Supramolecule #1: Single CMG purified from replicating Xenopus egg extracts

SupramoleculeName: Single CMG purified from replicating Xenopus egg extracts
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Xenopus laevis (African clawed frog)
Molecular weightTheoretical: 700 KDa

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Macromolecule #1: DNA replication licensing factor Mcm2 - Xenopus laevis

MacromoleculeName: DNA replication licensing factor Mcm2 - Xenopus laevis
type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MADSSESFNI ATSPRAGSRR DALTSSPGRD LPPFEDESEG MFGDGVVPEE EEDGEELIGD AMERDYRPI SELDRYEVEG LDDEEDVEDL TASQREAAEQ SMRMRDREMG RELGRMRRGL L YDSDEEEE DRPARKRRMA ERAAEGAPEE DEEMIESIEN LEDMKGHTVR ...String:
MADSSESFNI ATSPRAGSRR DALTSSPGRD LPPFEDESEG MFGDGVVPEE EEDGEELIGD AMERDYRPI SELDRYEVEG LDDEEDVEDL TASQREAAEQ SMRMRDREMG RELGRMRRGL L YDSDEEEE DRPARKRRMA ERAAEGAPEE DEEMIESIEN LEDMKGHTVR EWVSMAATRL EI YHRFKNF LRTHVDEHGH NVFKEKISDM CKENKESLPV NYEDLAAREH VLAYFLPEAP AEM LKIFDE AAKEVVLVMY PKYDRIAREI HVRISHLPLV EELRSLRQLH LNQLIRTSGV VTCC TGVLP QLSMVKYNCN KCNFILGPFF QSQNQEVRPG SCPECQSFGP FEINMEETVY QNYQR ITIQ ESPGKVAAGR LPRSKDAILL ADLVDSCKPG DEIELTGIYH NNYDGSLNTA NGFPVF ATV ILANHITKKD DKVAVGELTD EDVKAIVALS KDERIGERIF ASIAPSIYGH EDIKRGL AL ALFGGEAKNP GGKHKVRGDI NVLLCGDPGT AKSQFLKYVE KVASRAVFTT GQGASAVG L TAYVQRHPVT KEWTLEAGAL VLADRGVCLI DEFDKMNDQD RTSIHEAMEQ QSISISKAG IVTSLQARCT VIAASNPIGG RYDPSLTFSE NVDLTEPIVS RFDILCVVRD TVDPVQDEML ARFVVSSHI KHHPSSKDIA NGDAAEFALP NTFGVEALPQ EVLKKYIMYA KEKIRPKLNQ M DQDKVAKM YSDLRKESMA TGSIPITVRH IESMIRMAEA HARMHLRDYV VEDDVNMAIR VM LESFIDT QKFSVMRSMR KTFARYLAFR RDNNELLLFV LKQLIAEQVT YQRNRYGAQQ DTI EVPEKD LVDKARQINI HNLSAFYDSD LFKMNKFTHD VKKKLIIQQF

UniProtKB: DNA replication licensing factor mcm2

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Macromolecule #2: DNA replication licensing factor Mcm3 - Xenopus laevis

MacromoleculeName: DNA replication licensing factor Mcm3 - Xenopus laevis
type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MDYGGGFEDH ELREAQREYL DFLDDDQDQG LYHGKVRDMI GSNEHRLIVN LNDVRRKNDK RANLMLNDA FAETIAFQRA LKDLVASIDA TYAKQFEEFS VGFEGSFGSK HVSPRTLTAS L LGSLVCVE GIVTKCSLVR PKVMRSVHYC PATKKTLERK YSDLTSLEAF ...String:
MDYGGGFEDH ELREAQREYL DFLDDDQDQG LYHGKVRDMI GSNEHRLIVN LNDVRRKNDK RANLMLNDA FAETIAFQRA LKDLVASIDA TYAKQFEEFS VGFEGSFGSK HVSPRTLTAS L LGSLVCVE GIVTKCSLVR PKVMRSVHYC PATKKTLERK YSDLTSLEAF PSSSIYPTKD EE NNPLETE YGLSTYKDHQ TLSIQEMPEK APAGQLPRSV DIIADDDLVD KCKPGDRVQI VGI YRCLPS KQGGFTSGTF RTILLANNIK LMSKEIAPTF SADDVAKIKK FCKAHSKDIF EHLS KSLAP SIHGHEYIKK AILCMLLGGN EKVLENGTRI RGDINVLLIG DPSVAKSQLL RYVLH TAPR AIPTTGRGSS GVGLTAAVTT DQETGERRLE AGAMVLADRG VVCIDEFDKM SDMDRT AIH EVMEQGRVTI AKAGIQARLN ARCSVLAAAN PVYGRYDQYR TPMENIGLQD SLLSRFD LL FIVLDKMDAD NDQEIADHVL RMHRYRTPGE QDGYALPLGC SVEIFATDDP NASDVTDQ E LQIYEKHDNL LHGPRKNKSK IVSMQFIRKY IHVAKLIKPV LTSEAADYIS QEYAKIRNH DQINNDSART MPVTARALET MIRLSTAHAK VRMSKTIERQ DAETALELVQ FAYFKKVLAK EKKKTDKDL HDENLSQDTL SQESVRKSSR RAGKIADSQD DSMDPYSFSE QDSSLNENLS Q SLRPQRKK AESQDGKRSL SQNRTKEFKA ALLKAFKSSR SQSVAVSQLL ELINKGNPEP FE RSEVKEA LDNMQNDNQV MVSEDVVFLI

UniProtKB: Maternal DNA replication licensing factor mcm3

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Macromolecule #3: DNA replication licensing factor Mcm4 - Xenopus laevis

MacromoleculeName: DNA replication licensing factor Mcm4 - Xenopus laevis
type: protein_or_peptide / ID: 3 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MSSPTSTPSR RRNKRGRGSN PPTPHGEEVQ SPPSQRRRTE DSTSIGELLP MPTSPSGDVQ SPSGQELLF SSPAPSRHSA HQSELDLSSP LTYGTPSSRV EGTPRSGIRG TPARQRPDLG S ARKVKQVD LHSDQPAAEE LVTSEQSLGQ KLVIWGTDVN VATCKEKFQR ...String:
MSSPTSTPSR RRNKRGRGSN PPTPHGEEVQ SPPSQRRRTE DSTSIGELLP MPTSPSGDVQ SPSGQELLF SSPAPSRHSA HQSELDLSSP LTYGTPSSRV EGTPRSGIRG TPARQRPDLG S ARKVKQVD LHSDQPAAEE LVTSEQSLGQ KLVIWGTDVN VATCKEKFQR FVQRFIDPSA KE EDNVGLD LNEPIYMQRL EEINVVGDPF LNIDCDHLRN FDQDLYRQLV CYPQEVIPTF DMA ANEIFF ERYPDSILEH QIQVRPYNAL KTRNMRSLNP EDIDQLITIS GMVIRTSQII PEMQ EAFFK CQVCAFTTRV EIDRGRIAEP SVCKHCNTTH SMALIHNRSM FSDKQMIKLQ ESPED MPAG QTPHTTILYG HNDLVDKVQP GDRVNVTGIY RAVPIRVNPR VRNVKSVYKT HIDVIH YRK TDSKRLHGID EDTEQKLFTE ERVAMLKELA AKPDIYERLA AALAPSIYEH EDIKKGI LL QLFGGTRKDF SHTGRGKFRA EVNILLCGDP GTSKSQLLQY VFNLVPRGQY TSGKGSSA V GLTAYVMKDP ETRQLVLQTG ALVLSDNGIC CIDEFDKMNE STRSVLHEVM EQQTLSIAK AGIICQLNAR TSVLAAANPV ESQWNPKKTT IENIQLPHTL LSRFDLIFLM LDPQDEAYDR RLAHHLVAL YYQSEEQMKE EHLDMAVLKD YIAYARTYVN PRLSEEASQA LIEAYVSMRK I GSGRGMVS AYPRQLESLI RLSEAHAKVR FSNKVETIDV EEAKRLHREA LKQSATDPRT GI VDISILT TGMSATARKR KEELAQVLKK LIQSKGKTPA LKYQQLFEDL RGQSDAAITK DMF DEALHA LADDDYLTVT GKTVRLL

UniProtKB: DNA replication licensing factor mcm4-B

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Macromolecule #4: DNA replication licensing factor Mcm5 - Xenopus laevis

MacromoleculeName: DNA replication licensing factor Mcm5 - Xenopus laevis
type: protein_or_peptide / ID: 4 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MSGFDDLGVY YSDSFGGEQQ VGDDGQAKKS QLKKRFREFL RQYRIGTDRT GFTFKYRDEL KRHYNLGEY WIEVEMEDLA SFDEDLADYL YKQPTEHLQL LEEAAQEVAD EVTRPRPAGE E TIQEIQVM LRSDANPANI RSLKSEQMSH LVKIPGIIIA ATAVRAKATK ...String:
MSGFDDLGVY YSDSFGGEQQ VGDDGQAKKS QLKKRFREFL RQYRIGTDRT GFTFKYRDEL KRHYNLGEY WIEVEMEDLA SFDEDLADYL YKQPTEHLQL LEEAAQEVAD EVTRPRPAGE E TIQEIQVM LRSDANPANI RSLKSEQMSH LVKIPGIIIA ATAVRAKATK ISIQCRSCRN TI GNIAVRP GLEGYAMPRK CNTEQAGRPN CPLDPYFIIP DKCKCVDFQT LKLQESPDAV PHG ELPRHM QLYCDRYLCD KVVPGNRVTI MGIYSIRKSG KTSTKGRDRV GVGIRSSYIR VVGI QVDTE GTGRSAAGAI TPQEEEEFRR LAAKPDIYET VAKSIAPSIY GSSDIKKAIA CLLFG GSRK RLPDGLTRRG DVNLLMLGDP GTAKSQLLKF VERCSPIGVY TSGKGSSAAG LTASVM RDP VSRNFIMEGG AMVLADGGVV CIDEFDKMRE DDRVAIHEAM EQQTISIAKA GITTTLN SR CSVLAAANSV YGRWDDTKGE ENIDFMPTIL SRFDMIFIVK DEHNEQRDMT LAKHVMNV H LSARTQSSSV EGEVDLNTLK KYIAYCRAKC GPRLSAEAAE KLKNRYILMR SGAREHERE TEKRSSIPIT VRQLEAIVRI SESLGKMKLQ PFATETDVEE ALRLFQVSTL DAAMSGSLSG VEGFTTQED QEMLSRIEKQ MKKRFAIGSQ VSEHSIIQDF LKQKYPEHAI HKVLSLMMRR G EIQHRLQR KVLYRIK

UniProtKB: DNA replication licensing factor mcm5-A

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Macromolecule #5: DNA replication licensing factor Mcm6 - Xenopus laevis

MacromoleculeName: DNA replication licensing factor Mcm6 - Xenopus laevis
type: protein_or_peptide / ID: 5 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MELGGPAAAG DTDIAGQQLF KDELSDKCQK LFLEFLEECK GKDGSNLYVS AAEELIRPER NTLAVNFTD IEYYNQQLAT TIQEEYYRVY PHLCRAVRSF ARQMGNIPAN KEFYIAFSDF P ARQKIREL SSAKIGTLLR ISGQVVRTHP VHPELVSGTF LCMDCQSIVK ...String:
MELGGPAAAG DTDIAGQQLF KDELSDKCQK LFLEFLEECK GKDGSNLYVS AAEELIRPER NTLAVNFTD IEYYNQQLAT TIQEEYYRVY PHLCRAVRSF ARQMGNIPAN KEFYIAFSDF P ARQKIREL SSAKIGTLLR ISGQVVRTHP VHPELVSGTF LCMDCQSIVK DVEQQFRYTQ PT ICKNPVC ANRRRFTLDT NKSRFVDFQK VRIQETQAEL PRGAIPRSVE IILRAEAVES AMA GDRCDF TGTLIVVPDV SALAAGDARM ETGAKVTGGE GFNSEGVQGL KALGVRDLSY RLAF LACYV GATNPRFGGK DLREEDQTAE SIKNQMTVQE WEKVFEMSQD KNLYHNLCTS LFPTI HGND EIKRGVLLML FGGVPKTTME GTSLRGDINV CIVGDPSTSK SQFLKHVEEF SPRAVY TSG KASSAAGLTA AVVKDEESHE FVIEAGALML ADNGVCCIDE FDKMDLKDQV AIHEAME QQ TISITKAGVK ATLNARTSIL AAANPVGGRY ERSKSLKHNV NLSAPIMSRF DLFFILVD E CNEVTDYAIA RRIVDLHARN EESIERVYSI EDIQRYLLFA RQFQPKITKE AEEFIVEQY RRLRQRDGSG VAKSSWRITV RQLESLIRLS ESMARMHCSD EVQPKHVKEA FRLLSKSIIR VDTPDVSFD QGEDEKNIEG ENNGNLNNGE EAMETNQDEP INEKPSSNAG LKMSFAEYKQ I SNLLVLYM QKMEETEEEC HLTTTDLVNW YLKEMEAEIE TETELILKKR LIEKVIHRLI YY DHILIEL NKSELKTMDD TKETGEDAAE DRILVVNPNY MLED

UniProtKB: Maternal DNA replication licensing factor mcm6

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Macromolecule #6: DNA replication licensing factor Mcm7 - Xenopus laevis

MacromoleculeName: DNA replication licensing factor Mcm7 - Xenopus laevis
type: protein_or_peptide / ID: 6 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MPRDYQAEKE KCKTFLQEFY KDDEFGKKNF KYGVQLANIA HREQVALCID LDDLAEEDPE LVDAICENT RRYTNLFADA VQELLPQYKE REVVHKDALD VYIEHRLMME QRGRDPNEMR D PHNQYPPE LMRRFELYFK APSSSKARVV RDVKADSIGK LVTVRGIVTR ...String:
MPRDYQAEKE KCKTFLQEFY KDDEFGKKNF KYGVQLANIA HREQVALCID LDDLAEEDPE LVDAICENT RRYTNLFADA VQELLPQYKE REVVHKDALD VYIEHRLMME QRGRDPNEMR D PHNQYPPE LMRRFELYFK APSSSKARVV RDVKADSIGK LVTVRGIVTR VTEVKPMMVV AT YTCDQCG AETYQPIQSP TFMPLIMCPS RECQTNRSGG RLYLQTRGSK FIKFQELKIQ EHS DQVPVG NIPRCMSVYV RGENTRLAQP GDHVGITGVF LPMLRTGFRQ VVQGLLSETY LESH RLVKM NKTEDDELGT EELSEEELRQ ITEEDFYEKL AASIAPEIYG HEDVKKALLL LLVGG VDHS PRGMKIRGNI NVCLMGDPGV AKSQLLSYID RLAPRSQYTT GRGSSGVGLT AAVMKD PVT GEMTLEGGAL VLADQGVCCI DEFDKMMDSD RTAIHEVMEQ QTISIAKAGI MTTLNAR CS ILAAANPAYG RYNPKKTVEQ NIQLPAALLS RFDLLWLIQD KPDRDNDLRL AQHITYVH Q HSKQPPSQFQ PMDMKLMRRY ITMCKSKQPA IPESLADYLT AAYVEMRKEA RTNKDMTFT SARTLLSILR LSTALARLRL EDVVEKEDVN EAMRLTEMSK DSLQGDKGHA SRTQRPADVI FSTIREMVP EKGARSVKYS EAEQRCVSKG FTPAQFEAAL EEYEELNVWL VNQARTKITF V

UniProtKB: DNA replication licensing factor mcm7-B

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Macromolecule #7: DNA replication complex GINS protein Psf1 - Xenopus laevis

MacromoleculeName: DNA replication complex GINS protein Psf1 - Xenopus laevis
type: protein_or_peptide / ID: 7 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MFCEKAIELI RELQRASDGQ LPAFNEDGIR QILEEMKALY EQNQADVNEA KTEGRSDLIP TIKFRHCCL LRNRRCIVAY LYDRLLRIRA LRWEYGSVLP SALRFHMSTE EMDWFNQYKR S LATYMRSL GGEEGLDITQ DMKPPKSLYI EVRCLRDYGE FEIDDGTTIL ...String:
MFCEKAIELI RELQRASDGQ LPAFNEDGIR QILEEMKALY EQNQADVNEA KTEGRSDLIP TIKFRHCCL LRNRRCIVAY LYDRLLRIRA LRWEYGSVLP SALRFHMSTE EMDWFNQYKR S LATYMRSL GGEEGLDITQ DMKPPKSLYI EVRCLRDYGE FEIDDGTTIL LKKNSQHFLP RW KCEQLIR QGVLEHVLS

UniProtKB: DNA replication complex GINS protein PSF1

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Macromolecule #8: DNA replication complex GINS protein Psf2 - Xenopus laevis

MacromoleculeName: DNA replication complex GINS protein Psf2 - Xenopus laevis
type: protein_or_peptide / ID: 8 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString:
MDASEVEFLA EKEQVTVIPN FSLDKVYLIG GDLGPFNPSL PVEVPLWLAI NLKQRQKCRI VPPEWMDVE KLEAIRDQER REETFTPMPS PYYMELTKLL LNHAADNIPK ADEIRTLVKD T WDTRIAKL RLSADSFVKG QEAHAKLDNL TLMEINTIGT FFTESLNHMY KLRTSLQNPE EG QSQDY

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Macromolecule #9: DNA replication complex GINS protein Psf3 - Xenopus laevis

MacromoleculeName: DNA replication complex GINS protein Psf3 - Xenopus laevis
type: protein_or_peptide / ID: 9 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MWEPYMPVEP GLGREENFLS LEDLLMSQEK LPCCIESGFP RLGFLDKGGD SDSIPEGSKM ELPLWLAKG LYDNKRRVLS VELPKIYREG WRTVFSADAN VVDLHKMGPH YYGFGSQLLN F DSPENPEI AKTILQTFVG RFRRIMDSSQ NAYNEDTSGL VARLDELERS ...String:
MWEPYMPVEP GLGREENFLS LEDLLMSQEK LPCCIESGFP RLGFLDKGGD SDSIPEGSKM ELPLWLAKG LYDNKRRVLS VELPKIYREG WRTVFSADAN VVDLHKMGPH YYGFGSQLLN F DSPENPEI AKTILQTFVG RFRRIMDSSQ NAYNEDTSGL VARLDELERS LFRAGQRGLN AF QSWERGK AAQITASNLV QNYKKRKFNE ADA

UniProtKB: DNA replication complex GINS protein PSF3

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Macromolecule #10: DNA replication complex GINS protein Sld5 - Xenopus laevis

MacromoleculeName: DNA replication complex GINS protein Sld5 - Xenopus laevis
type: protein_or_peptide / ID: 10 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MEDELALSDQ GSDEDEEVLT PAELINKLEE AWLNEKFAPE LLESKSEVVE CVMEQLNHME QNLHRAKPG DLKISFHHME IERIRYMLSS YLRSRMLKIE KFFPHILEKE KSRGEGEPPH L SPEEFAFA KEYMTNTETL LKSVALRHMP PNLQTVDLLK SVPKPNLDSF ...String:
MEDELALSDQ GSDEDEEVLT PAELINKLEE AWLNEKFAPE LLESKSEVVE CVMEQLNHME QNLHRAKPG DLKISFHHME IERIRYMLSS YLRSRMLKIE KFFPHILEKE KSRGEGEPPH L SPEEFAFA KEYMTNTETL LKSVALRHMP PNLQTVDLLK SVPKPNLDSF VFLRVKEEQN NI LVEPETD EQSEYAIDME VGSQHLIRYR TIAPLVASGA VKLI

UniProtKB: DNA replication complex GINS protein SLD5

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Macromolecule #11: Cell division control protein 45 - Xenopus laevis

MacromoleculeName: Cell division control protein 45 - Xenopus laevis / type: protein_or_peptide / ID: 11 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
SequenceString: MFVSDLRKEF FDVIVTERVL LLVAPDVDAL CACKILQALF QCDHVQYTLV PVSGWQELET LFLEHKEQF RYFVLINCGA NIDLLETLQP QEEAIFYICD THRPIDVVNI YNDSQVKLLI R QDDDLEIP AYDDIFNDDE EDGEDSGNES DGAEPSGKRR RFDEAAVERR ...String:
MFVSDLRKEF FDVIVTERVL LLVAPDVDAL CACKILQALF QCDHVQYTLV PVSGWQELET LFLEHKEQF RYFVLINCGA NIDLLETLQP QEEAIFYICD THRPIDVVNI YNDSQVKLLI R QDDDLEIP AYDDIFNDDE EDGEDSGNES DGAEPSGKRR RFDEAAVERR IERRRQRREW EA RRREIIF DYEQYEYHGT SSAMMMFELA WIMSKDSNDM LWWAIVGLTD QWVQDRITQM KYV TDVGTL QRHVSRHNHR NEDEENSLSI DCMRIAFEYD LRLSLYQHWS LYESICNSCY TSAT LKLWS LQGQKKLQEF LADMGMPLKQ VKQKFNSMDI SLKENLREML EESANKFGMK DVRVQ TFSV QFGFKNKFLA SDIVFAVLSL LENTERDEKG TDNFIKALDS LSRSNLDKLH TGLEMG KKL LCAIQQTVAS CICTNLILSQ GPFLYCYLME GTPDVKMFSN PISLCLLCKY LLKSFVC ST KNKRCKLLPL VLAAPLDAEK GTVIMVGIPP EAESSDKKNF FGRAFEKAAE STSSRTLH N HFDMSIIELR TEDRSKFLDA LISLLS

UniProtKB: Cell division control protein 45 homolog

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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: 7.5
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.4 µm / Nominal defocus min: 1.3 µm
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 33.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: PROJECTION MATCHING
Final angle assignmentType: PROJECTION MATCHING
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.03 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 137591

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