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- EMDB-17460: S. cerevisiae sCMGE with N-ter Mcm10 density -

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Basic information

Entry
Database: EMDB / ID: EMD-17460
TitleS. cerevisiae sCMGE with N-ter Mcm10 density
Map dataEM map of sCMG10
Sample
  • Complex: Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsilon (subunit A and B), 19-mer DNA molecule with complementary 7-mer DNA molecule
    • Complex: DNA replication complex GINS containing Psf1, Psf2, Psf3 and Sld5
      • Protein or peptide: x 2 types
      • DNA: x 2 types
    • Complex: DNA replication licensing factors MCM2-7
      • Protein or peptide: x 6 types
    • Complex: DNA polymerase epsilon contains catalytic subunit A and non-catalytic subunit B
      • Protein or peptide: x 2 types
    • Complex: DNA
      • Protein or peptide: x 1 types
    • Complex: Cdc45
      • Protein or peptide: x 1 types
KeywordsSaccharomyces cerevisiae / helicase / CMGE / initiation of DNA replication / DNA / DNA unwinding / REPLICATION / Mcm10
Biological speciesSaccharomyces cerevisiae (brewer's yeast) / DNA molecule (others)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.5 Å
AuthorsHenrikus SS / Willhoft O
Funding support United Kingdom, European Union, France, 4 items
OrganizationGrant numberCountry
The Francis Crick InstituteFC001065 and FC001066 United Kingdom
European Research Council (ERC)grant agreement no. 820102European Union
Human Frontier Science Program (HFSP)LT000834/2020-L France
European Molecular Biology Organization (EMBO)ALTF 962-2019European Union
CitationJournal: Nat Struct Mol Biol / Year: 2024
Title: Unwinding of a eukaryotic origin of replication visualized by cryo-EM.
Authors: Sarah S Henrikus / Marta H Gross / Oliver Willhoft / Thomas Pühringer / Jacob S Lewis / Allison W McClure / Julia F Greiwe / Giacomo Palm / Andrea Nans / John F X Diffley / Alessandro Costa /
Abstract: To prevent detrimental chromosome re-replication, DNA loading of a double hexamer of the minichromosome maintenance (MCM) replicative helicase is temporally separated from DNA unwinding. Upon S-phase ...To prevent detrimental chromosome re-replication, DNA loading of a double hexamer of the minichromosome maintenance (MCM) replicative helicase is temporally separated from DNA unwinding. Upon S-phase transition in yeast, DNA unwinding is achieved in two steps: limited opening of the double helix and topological separation of the two DNA strands. First, Cdc45, GINS and Polε engage MCM to assemble a double CMGE with two partially separated hexamers that nucleate DNA melting. In the second step, triggered by Mcm10, two CMGEs separate completely, eject the lagging-strand template and cross paths. To understand Mcm10 during helicase activation, we used biochemical reconstitution with cryogenic electron microscopy. We found that Mcm10 splits the double CMGE by engaging the N-terminal homo-dimerization face of MCM. To eject the lagging strand, DNA unwinding is started from the N-terminal side of MCM while the hexamer channel becomes too narrow to harbor duplex DNA.
History
DepositionMay 25, 2023-
Header (metadata) releaseMay 29, 2024-
Map releaseMay 29, 2024-
UpdateAug 28, 2024-
Current statusAug 28, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_17460.png

Downloads & links

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Map

FileDownload / File: emd_17460.map.gz / Format: CCP4 / Size: 22.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationEM map of sCMG10
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.16 Å/pix.
x 180 pix.
= 388.8 Å		2.16 Å/pix.
x 180 pix.
= 388.8 Å		2.16 Å/pix.
x 180 pix.
= 388.8 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.16 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.017
Minimum - Maximum-0.024492262 - 0.092643544
Average (Standard dev.)0.0005687723 (±0.0039453125)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions180180180
Spacing180180180
CellA=B=C: 388.80002 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: EM half map 2 of sCMG10

Fileemd_17460_half_map_1.map
AnnotationEM half map 2 of sCMG10
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsi...

EntireName: Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsilon (subunit A and B), 19-mer DNA molecule with complementary 7-mer DNA molecule
Components
  • Complex: Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsilon (subunit A and B), 19-mer DNA molecule with complementary 7-mer DNA molecule
    • Complex: DNA replication complex GINS containing Psf1, Psf2, Psf3 and Sld5
      • Protein or peptide: DNA replication complex GINS protein PSF1
      • Protein or peptide: DNA replication complex GINS protein PSF2
      • DNA: DNA strand 1
      • DNA: DNA strand 2
    • Complex: DNA replication licensing factors MCM2-7
      • Protein or peptide: DNA replication licensing factor MCM2
      • Protein or peptide: DNA replication licensing factor MCM3
      • Protein or peptide: DNA replication licensing factor MCM4
      • Protein or peptide: DNA replication licensing factor MCM5
      • Protein or peptide: DNA replication licensing factor MCM6
      • Protein or peptide: Cell division control protein 45
    • Complex: DNA polymerase epsilon contains catalytic subunit A and non-catalytic subunit B
      • Protein or peptide: DNA replication complex GINS protein SLD5
      • Protein or peptide: Minichromosome maintenance protein 10
    • Complex: DNA
      • Protein or peptide: DNA replication licensing factor MCM7
    • Complex: Cdc45
      • Protein or peptide: DNA replication complex GINS protein PSF3

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Supramolecule #1: Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsi...

SupramoleculeName: Complex of Mcm2-7, Cdc45, GINS (Psf1-3, Sld5) DNA polymerase epsilon (subunit A and B), 19-mer DNA molecule with complementary 7-mer DNA molecule
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#14
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Supramolecule #2: DNA replication complex GINS containing Psf1, Psf2, Psf3 and Sld5

SupramoleculeName: DNA replication complex GINS containing Psf1, Psf2, Psf3 and Sld5
type: complex / ID: 2 / Parent: 1 / Macromolecule list: #7-#8, #12-#13
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Supramolecule #3: DNA replication licensing factors MCM2-7

SupramoleculeName: DNA replication licensing factors MCM2-7 / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #1-#6
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Supramolecule #4: DNA polymerase epsilon contains catalytic subunit A and non-catal...

SupramoleculeName: DNA polymerase epsilon contains catalytic subunit A and non-catalytic subunit B
type: complex / ID: 4 / Parent: 1 / Macromolecule list: #10-#11
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Supramolecule #5: DNA

SupramoleculeName: DNA / type: complex / ID: 5 / Parent: 1 / Macromolecule list: #14
Source (natural)Organism: DNA molecule (others)

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Supramolecule #6: Cdc45

SupramoleculeName: Cdc45 / type: complex / ID: 6 / Parent: 1 / Macromolecule list: #9
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Macromolecule #1: DNA replication licensing factor MCM2

MacromoleculeName: DNA replication licensing factor MCM2 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: EELTLESLSN VKANSYSEWI TQPNVSRTIA RELKSFLLEY TDETGRSVYG ARIRTLGEMN SESLEVNYR HLAESKAILA LFLAKCPEEM LKIFDLVAME ATELHYPDYA RIHSEIHVRI S AAAAIYSL RELRESNLSS LVRVTGVVTR RTGVFPQLKY VKFNCLKCGS ...String:
EELTLESLSN VKANSYSEWI TQPNVSRTIA RELKSFLLEY TDETGRSVYG ARIRTLGEMN SESLEVNYR HLAESKAILA LFLAKCPEEM LKIFDLVAME ATELHYPDYA RIHSEIHVRI S AAAAIYSL RELRESNLSS LVRVTGVVTR RTGVFPQLKY VKFNCLKCGS ILGPFFQDSN EE IRISFCT NCKSKGPFRV NGEKTVYRNY QRVTLQEAPG TVPPGRLPRH REVILLADLV DVS KPGEEV EVTGIYKNNY DGNLNAKNGF PVFATIIEAN SIKRREGNTA NEGEEGLDVF SWTE EEERE FRKISRDRGI IDKIISSMAP SIYGHRDIKT AVACSLFGGV PKNVNGKHSI RGDIN VLLL GDPGTAKSQI LKYVEKTAHR AVFATGQGAS AVGLTASVRK DPITKEWTLE GGALVL ADK GVCLIDEFDK MNDQDRTSIH EAMEQQSISI SKAGIVTTLQ ARCSIIAAAN PNGGRYN ST LPLAQNVSLT EPILSRFDIL CVVRDLVDEE ADERLATFVV DSHVRSHPEN LNARQRRL Q RQRKKEEEIS PIPQELLMKY IHYARTKIYP KLHQMDMDKV SRVYADLRRE SISTGSFPI TVRHLESILR IAESFAKMRL SEFVSSYDLD RAIKVVVDSF VDAQKVSVRR QLRRSFAIYT LG

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Macromolecule #2: DNA replication licensing factor MCM3

MacromoleculeName: DNA replication licensing factor MCM3 / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: DAVFGDRVRR FQEFLDTFTS YRDSVRSIQV YNSNNAANYL NILPHRIIIS LDDLREFDRS FWSGILVEP AYFIPPAEKA LTDLADSMDD VPAPNASAVS SRHPWKLSFK GSFGAHALSP R TLTAQHLN KLVSVEGIVT KTSLVRPKLI RSVHYAAKTG RFHYRDYTDA ...String:
DAVFGDRVRR FQEFLDTFTS YRDSVRSIQV YNSNNAANYL NILPHRIIIS LDDLREFDRS FWSGILVEP AYFIPPAEKA LTDLADSMDD VPAPNASAVS SRHPWKLSFK GSFGAHALSP R TLTAQHLN KLVSVEGIVT KTSLVRPKLI RSVHYAAKTG RFHYRDYTDA TTTLTTRIPT PA IYPTEDT EGNKLTTEYG YSTFIDHQRI TVQEMPEMAP AGQLPRSIDV ILDDDLVDKT KPG DRVNVV GVFKSLGAGG MNQSNSNTLI GFKTLILGNT VYPLHARQML TDFDIRNINK LSKK KDIFD ILSQSLAPSI YGHDHIKKAI LLMLMGGVEK NLENGSHLRG DINILMVGDP STAKS QLLR FVLNTASLAI ATTGRGSSGV GLTAAVTTDR ETGERRLEAG AMVLADRGVV CIDEFD KMT DVDRVAIHEV MEQQTVTIAK AGIHTTLNAR CSVIAAANPV FGQYDVNRDP HQNIALP DS LLSRFDLLFV VTDDINEIRD RSISEHVLRT HRYLPPGYLE GEPVRERLNL SLAVGEQA G AKLAKNKGNY NGTEIPKLVT IPFLRKYVQY AKERVIPQLT QEAINVIVKN YTDLRNDDN TKKSPITART LETLIRLATA HAKVRLSKTV NKVDAKVAAN LLRFALLGE

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Macromolecule #3: DNA replication licensing factor MCM4

MacromoleculeName: DNA replication licensing factor MCM4 / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: TNVSIQECTT NFRNFLMSFK YKFRKILDER ELYYIKQLNE MRELGTSNLN LDARNLLAYK QTEDLYHQL LNYPQEVISI MDQTIKDCMV SLIVDLDEIE TKFYKVRPYN VGSCKGMREL N PNDIDKLI NLKGLVLRST PVIPDMKVAF FKCNVCDHTM AVEIDRGVIQ ...String:
TNVSIQECTT NFRNFLMSFK YKFRKILDER ELYYIKQLNE MRELGTSNLN LDARNLLAYK QTEDLYHQL LNYPQEVISI MDQTIKDCMV SLIVDLDEIE TKFYKVRPYN VGSCKGMREL N PNDIDKLI NLKGLVLRST PVIPDMKVAF FKCNVCDHTM AVEIDRGVIQ EPARCERIDC NE PNSMSLI HNRCSFADKQ VIKLQETPDF VPDGQTPHSI SLCVYDELVD SCRAGDRIEV TGT FRSIPI RANSRQRVLK SLYKTYVDVV HVKKVQDLAK IREVAAREDL YSLLARSIAP SIYE LEDVK KGILLQLFGG TNKTFTKGGR YRGDINILLC GDPSTSKSQI LQYVHKITPR GVYTS GGLT AYITRDVDTK QLVLESGALV LSDGGVCCID EFDKMSDSTR SVLHEVMEQQ TISIAK AGI ITTLNARSSI LASANPIGSR YNPNLPVTEN IDLPPPLLSR FDLVYLVLDK VDEKNDR EL AKHLTNLYLV LPVEFLTMYI SYAKEHIHPI ITEAAKTELV RAYVGMRKMG DDSRSDEK R ITATTRQLES MIRLAEAHAK MKLKNVVELE DVQEAVRLIR SAIKDYATDP K

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Macromolecule #4: DNA replication licensing factor MCM5

MacromoleculeName: DNA replication licensing factor MCM5 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: NDDDNTEIIK SFKNFILEFR LDSQFIYRDQ LRNNILVKNY SLTVNMEHLI GYNEDIYKKL SDEPSDIIP LFETAITQVA KRISILSRAQ ALNSLPTFQL ILNSNANQIP LRDLDSEHVS K IVRLSGII ISTSVLSSRA TYLSIMCRNC RHTTSITINN FNSIPNNTVS ...String:
NDDDNTEIIK SFKNFILEFR LDSQFIYRDQ LRNNILVKNY SLTVNMEHLI GYNEDIYKKL SDEPSDIIP LFETAITQVA KRISILSRAQ ALNSLPTFQL ILNSNANQIP LRDLDSEHVS K IVRLSGII ISTSVLSSRA TYLSIMCRNC RHTTSITINN FNSIPNNTVS LPRSCLSKNC GP DPYIIIH ESSKFIDQQF LKLQEIPELV PVGEMPRNLT MTCDRYLTNK VIPGTRVTIV GIY SIYNSK NASGVAIRTP YIKILGIQSD VETSSINSVT MFTEEEEEEF LQLSRNPKLY EILT NSIAP SIFGNEDIKK AIVCLLMGGS KKILPDGMRL RGDINVLLLG DPGTAKSQLL KFVEK VSPI AVYTSGKGSS AAGLTASVQR DPMTREFYLE GGAMVLADGG VVCIDEFDKM RDEDRV AIH EAMEQQTISI AKAGITTVLN SRTSVLAAAN PIYGRYDDLK SPGDNIDFQT TILSRFD MI FIVKDDHNEE RDISIANHVI NIHTGNANAM ENQQEENGSE ISIEKMKRYI TYCRLKCA P RLSPQAAEKL SSNFVTIRKQ LLINELESTE RSSIPITIRQ LEAIIRITES LAKLELSPI AQERHVDEAI RLFQASTMDA ASQDPIGGTS LSEIRRFEQE LKRRLPIGWS TSYQTLRREF VDTHAFSQL ALDKALYALA AETIQLRHQG QNIYRS

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Macromolecule #5: DNA replication licensing factor MCM6

MacromoleculeName: DNA replication licensing factor MCM6 / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: LNHVKKVDDV TGEKVREAFE QFLEDFSVQS TDTGEVEKVY RAQIEFMKIY DLNTIYIDYQ HLSMRENGA LAMAISEQYY RFLPFLQKGL RRVVRKYAPE LLNERVFQIS FFNLPTVHRI R DIRSEKIG SLLSISGTVT RTSEVRPELY KASFTCDMCR AIVDNVEQSF ...String:
LNHVKKVDDV TGEKVREAFE QFLEDFSVQS TDTGEVEKVY RAQIEFMKIY DLNTIYIDYQ HLSMRENGA LAMAISEQYY RFLPFLQKGL RRVVRKYAPE LLNERVFQIS FFNLPTVHRI R DIRSEKIG SLLSISGTVT RTSEVRPELY KASFTCDMCR AIVDNVEQSF KYTEPTFCPN PS CENRAFW TLNVTRSRFL DWQKVRIQEN ANEIPTGSMP RTLDVILRGD SVERAKPGDR CKF TGVEIV VPDVTQLGLP GVKKTTEGLN SGVTGLRSLG VRDLTYKISF LACHVISIGD QEVF LNSLS SDEINELKEM VKDEHIYDKL VRSIAPAVFG HEAVKKGILL QMLGGVHKST VEGIK LRGD INICVVGDPS TSKSQFLKYV VGFAPRSVYT SGKASSAAGL TAAVVRDEEG GDYTIE AGA LMLADNGICC IDEFDKMDIS DQVAIHEAME QQTISIAKAG IHATLNARTS ILAAANP VG GRYNRKLSLR GNLNMTAPIM SRFDLFFVIL DDCNEKIDTE LASHIVDLHM KRDEAIEP P FSAEQLRRYI KYARTFKPIL TKEARSYLVE KYKELRKDDA QGFSRSSYRI TVRQLESMI RLSEAIARAN CVDEITPSFI AEAYDLLRQS IIRVDV

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Macromolecule #6: Cell division control protein 45

MacromoleculeName: Cell division control protein 45 / type: protein_or_peptide / ID: 6 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MYYGISQFSE AYNKILRNSS SHSSCQLVIF VSCLNIDALC ATKMLSLLFK KQLVQSQIVP IFGYSELRR HYSQLDDNIN SLLLVGFGGV IDLEAFLEID PQEYVISFRR DIYVLDAHRP W NLDNIFGS QIIQCFDDGT VDDTLGEQKE AYYKLLRKQR KKQIHEYEGV ...String:
MYYGISQFSE AYNKILRNSS SHSSCQLVIF VSCLNIDALC ATKMLSLLFK KQLVQSQIVP IFGYSELRR HYSQLDDNIN SLLLVGFGGV IDLEAFLEID PQEYVISFRR DIYVLDAHRP W NLDNIFGS QIIQCFDDGT VDDTLGEQKE AYYKLLRKQR KKQIHEYEGV LEEYYSQGTT VV NSISAQI YSLLSAIGET NLSNLWLNIL GTTSLDIAYA QVYNRLYPLL QDEVKRLVKT PDT LTLNIQ PDYYLFLLRH SSLYDSFYYS NYVNAKLSLW NENGKKRLHK MFARMGIPLS TAQE TWLYM DHSIKRELGI IFDKNLDRYG LQDIIRDGFV RTLGYRGSIS ASEFVEALTA LLEVG NSAQ KLTNLRKRWV SNFWLSWDAL DDRKVELLNR GIQLAQDLQR AIFNTGVAIL EKKLIK HLR IYRLCVLQDG PDLDLYRNPL TLLRLGNWLI ECCAESEDKQ LLPMVLASID ENTDTYL VA GLTPRYPRGL DTIHTKKPIL NNFSMAFQQI TAETDAKVRI DNFESSIIEI RREDLSPF L EKLTLSGL

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Macromolecule #7: DNA replication complex GINS protein PSF1

MacromoleculeName: DNA replication complex GINS protein PSF1 / type: protein_or_peptide / ID: 7 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: YGDLGNKLVL EAKRTKQLYA RSNQDVNLPM YHEDIIRNIL KEVSNLRKNT EYLKEQQQLG MLDDKVAKC QYFVTLLCME RNKRCLLAYQ RLRTDILDSM AWNNNGLDLM SSSQQDTNNL S HQEQEYLK EYCDLITDLK SGDLVDIDLS GSLVPPSDVF IDVRVLKDAG ...String:
YGDLGNKLVL EAKRTKQLYA RSNQDVNLPM YHEDIIRNIL KEVSNLRKNT EYLKEQQQLG MLDDKVAKC QYFVTLLCME RNKRCLLAYQ RLRTDILDSM AWNNNGLDLM SSSQQDTNNL S HQEQEYLK EYCDLITDLK SGDLVDIDLS GSLVPPSDVF IDVRVLKDAG EIQTEYGVFN LI KDSQFFV RQSDVERLIQ QGYLQKI

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Macromolecule #8: DNA replication complex GINS protein PSF2

MacromoleculeName: DNA replication complex GINS protein PSF2 / type: protein_or_peptide / ID: 8 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString:
SLPAHLQQTF SPEEIQFIVE NEPIKIFPRI TTRQKIHTRW QLITTDDKAL NNMVAMRSTE VVLWIALLL KQQSKCSIVA PQWLTTKELD RKIQYEKTHP DRFSELPWNW LVLARILFNK A KDDFHDPI HELRGKIQDL REIRQIKVLK GLKYLNESHL QLDNLSLLEI NELRPFITEI MD KLREIHT ASL

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Macromolecule #9: DNA replication complex GINS protein PSF3

MacromoleculeName: DNA replication complex GINS protein PSF3 / type: protein_or_peptide / ID: 9 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString:
MGYYDIDDVL ADGTEFPCKF QYDIPGLGYL ENNPGRPITK NTKLSLPLWL ARILAIVGGD EPVPFVELL PPDMFSTKVM NAIKTDPVAL DLHSINSHFF SLAIKWIMLF SEKELANVVS E LLLQRAQE LNHHASSLSI NTNIATSTFL LKLEEMEKEI YKKSHESYKD TKRWMFK

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Macromolecule #10: DNA replication complex GINS protein SLD5

MacromoleculeName: DNA replication complex GINS protein SLD5 / type: protein_or_peptide / ID: 10 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: TQIYVSPQQD FSDLMKSWKN ERCSPELLPY PHQLMKRLLN RISMQSQLIE NISMGFLDMQ NASNANPPM PNESKLPLLC METELERLKF VIRSYIRCRL SKIDKFSLYL RQLNEDENSL I SLTDLLSK DEIKYHDTHS LIWLKLVNDS ILKYMPEELQ AINDTEGSVN ...String:
TQIYVSPQQD FSDLMKSWKN ERCSPELLPY PHQLMKRLLN RISMQSQLIE NISMGFLDMQ NASNANPPM PNESKLPLLC METELERLKF VIRSYIRCRL SKIDKFSLYL RQLNEDENSL I SLTDLLSK DEIKYHDTHS LIWLKLVNDS ILKYMPEELQ AINDTEGSVN MIDEPDWNKF VF IHVNGPP DGKWNEDPLL QENEFGKPCY TVTIPDLKEE VELTIGSIYV MRYEVIRDLL RDD KVALI

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Macromolecule #11: Minichromosome maintenance protein 10

MacromoleculeName: Minichromosome maintenance protein 10 / type: protein_or_peptide / ID: 11 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MNDPREILAV DPYNNITSDE EDEQAIAREL EFMERKRQAL VERLKRKQEF KKPQDPNFEA IEVPQSPTK NRVKVGSHNA TQQGTKFEGS NINEVRLSQL QQQPKPPAST TTYFMEKFQN A KKNEDKQI AKFESMMNAR VHTFSTDEKK YVPIITNELE SFSNLWVKKR ...String:
MNDPREILAV DPYNNITSDE EDEQAIAREL EFMERKRQAL VERLKRKQEF KKPQDPNFEA IEVPQSPTK NRVKVGSHNA TQQGTKFEGS NINEVRLSQL QQQPKPPAST TTYFMEKFQN A KKNEDKQI AKFESMMNAR VHTFSTDEKK YVPIITNELE SFSNLWVKKR YIPEDDLKRA LH EIKILRL GKLFAKIRPP KFQEPEYANW ATVGLISHKS DIKFTSSEKP VKFFMFTITD FQH TLDVYI FGKKGVERYY NLRLGDVIAI LNPEVLPWRP SGRGNFIKSF NLRISHDFKC ILEI GSSRD LGWCPIVNKK THKKCGSPIN ISLHKCCDYH REVQFRGTSA KRIELNGGYA LGAPT KVDS QPSLYKAKGE NGFNIIKGTR KRLSEEEERL KKSSHNFTNS NSAKAFFDEK FQNPDM LAN LDNKRRKIIE TKKSTALSRE LGKIMRRRES SGLEDKSVGE RQKMKRTTES ALQTGLI QR LGFDPTHGKI SQVLKSSVSG SEPKNNLLGK KKTVINDLLH YKKEKVILAP SKNEWFKK R SHREEVWQKH FGSKETKETS DGSASDLEII

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Macromolecule #14: DNA replication licensing factor MCM7

MacromoleculeName: DNA replication licensing factor MCM7 / type: protein_or_peptide / ID: 14 / Enantiomer: LEVO
Source (natural)Organism: DNA molecule (others)
Recombinant expressionOrganism: DNA molecule (others)
SequenceString: ALPSIQLPVD YNNLFNEITD FLVTFKQKGP KYMAMLQKVA NRELNSVIID LDDILQYQNE KFLQGTQAD DLVSAIQQNA NHFTELFCRA IDNNMPLPTK EIDYKDDVLD VILNQRRLRN E RMLTELFP PNLTRRYFLY FKPLSQNCAR RYRKKAISSK PLSVRQIKGD ...String:
ALPSIQLPVD YNNLFNEITD FLVTFKQKGP KYMAMLQKVA NRELNSVIID LDDILQYQNE KFLQGTQAD DLVSAIQQNA NHFTELFCRA IDNNMPLPTK EIDYKDDVLD VILNQRRLRN E RMLTELFP PNLTRRYFLY FKPLSQNCAR RYRKKAISSK PLSVRQIKGD FLGQLITVRG II TRVSDVK PAVEVIAYTC DQCGYEVFQE VNSRTFTPLS ECTSEECSQN QTKGQLFMST RAS KFSAFQ ECKIQELSQQ VPVGHIPRSL NIHVNGTLVR SLSPGDIVDV TGIFLPAPYT GFKA LKAGL LTETYLEAQF VRQHKKKFAS FSLTSDVEER VMELITSGDV YNRLAKSIAP EIYGN LDVK KALLLLLVGG VDKRVGDGMK IRGDINVCLM GDPGVAKSQL LKAICKISPR GVYTTG KGS SGVGLTAAVM KDPVTDEMIL EGGALVLADN GICCIDEFDK MDESDRTAIH EVMEQQT IS ISKAGINTTL NARTSILAAA NPLYGRYNPR LSPLDNINLP AALLSRFDIL FLMLDIPS R DDDEKLAEHV TYVHMHNKQP DLDFTPVEPS KMREYIAYAK TKRPVMSEAV NDYVVQAYI RLRQDSKREM DSKFSFGQAT PRTLLGIIRL SQALAKLRLA DMVDIDDVEE ALRLVRVSKE SLY

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Macromolecule #12: DNA strand 1

MacromoleculeName: DNA strand 1 / type: dna / ID: 12 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString:
AAAAAAAAAA AAAAAAAAA

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Macromolecule #13: DNA strand 2

MacromoleculeName: DNA strand 2 / type: dna / ID: 13 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString:
TTTTTTT

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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 / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 1.67 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.3000000000000003 µm / Nominal defocus min: 1.8 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: OTHER / Details: Ab initio reconstruction in cryoSPARC v4.0.3
Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.5 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 85107
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
FSC plot (resolution estimation)

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