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- EMDB-23755: CryoEM map of the structure of the S. cerevisiae origin recogniti... -

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Basic information

Entry
Database: EMDB / ID: EMD-23755
TitleCryoEM map of the structure of the S. cerevisiae origin recognition complex bound to the replication initiator Cdc6 and the ARS1 origin DNA.
Map data
Sample
  • Complex: Complex of origin replication complex with Cdc6 and ARS1 origin DNA
    • Protein or peptide: x 7 types
    • DNA: x 2 types
  • Ligand: x 2 types
Function / homology
Function and homology information


CDK-mediated phosphorylation and removal of Cdc6 / CDC6 association with the ORC:origin complex / Assembly of the ORC complex at the origin of replication / Orc1 removal from chromatin / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / nuclear origin of replication recognition complex / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / nuclear pre-replicative complex ...CDK-mediated phosphorylation and removal of Cdc6 / CDC6 association with the ORC:origin complex / Assembly of the ORC complex at the origin of replication / Orc1 removal from chromatin / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / nuclear origin of replication recognition complex / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / mitotic DNA replication checkpoint signaling / silent mating-type cassette heterochromatin formation / regulation of DNA-templated DNA replication initiation / DNA replication origin binding / regulation of DNA replication / subtelomeric heterochromatin formation / DNA replication initiation / nucleosome binding / G1/S transition of mitotic cell cycle / chromosome / chromosome, telomeric region / cell division / GTPase activity / chromatin binding / GTP binding / ATP hydrolysis activity / nucleoplasm / ATP binding / metal ion binding / nucleus / cytoplasm
Similarity search - Function
Origin recognition complex, subunit 6, fungi / Cell division protein Cdc6/18 / : / Origin recognition complex subunit 1 C-terminal winged HTH domain / Origin recognition complex, subunit 6 / Origin recognition complex subunit 6 (ORC6) / AAA domain / Origin recognition complex subunit 4 / Origin recognition complex, subunit 3 / Origin recognition complex, subunit 5 ...Origin recognition complex, subunit 6, fungi / Cell division protein Cdc6/18 / : / Origin recognition complex subunit 1 C-terminal winged HTH domain / Origin recognition complex, subunit 6 / Origin recognition complex subunit 6 (ORC6) / AAA domain / Origin recognition complex subunit 4 / Origin recognition complex, subunit 3 / Origin recognition complex, subunit 5 / Origin recognition complex subunit 4, C-terminal / Origin recognition complex subunit 3, winged helix C-terminal / : / : / Origin recognition complex (ORC) subunit 4 C-terminus / Origin recognition complex (ORC) subunit 5 C-terminus / Origin recognition complex winged helix C-terminal / ORC5, lid domain / Orc1-like, AAA ATPase domain / AAA ATPase domain / Origin recognition complex subunit 2 / Origin recognition complex, subunit 2 / AAA lid domain / AAA lid domain / Bromo adjacent homology (BAH) domain superfamily / Bromo adjacent homology domain / Bromo adjacent homology (BAH) domain / BAH domain / BAH domain profile. / ATPase family associated with various cellular activities (AAA) / ATPase, AAA-type, core / EF-Hand 1, calcium-binding site / Winged helix DNA-binding domain superfamily / Winged helix-like DNA-binding domain superfamily / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
Cell division control protein 6 / Origin recognition complex subunit 2 / Origin recognition complex subunit 6 / Origin recognition complex subunit 5 / Origin recognition complex subunit 1 / Origin recognition complex subunit 3 / Origin recognition complex subunit 4
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.6 Å
AuthorsFeng X / Li H
Funding support United States, 4 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM131754 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM45436 United States
Biotechnology and Biological Sciences Research Council (BBSRC)BB/S001387.1 United States
Biotechnology and Biological Sciences Research Council (BBSRC)BB/N000323/1 United States
CitationJournal: Nat Commun / Year: 2021
Title: The structure of ORC-Cdc6 on an origin DNA reveals the mechanism of ORC activation by the replication initiator Cdc6.
Authors: Xiang Feng / Yasunori Noguchi / Marta Barbon / Bruce Stillman / Christian Speck / Huilin Li /
Abstract: The Origin Recognition Complex (ORC) binds to sites in chromosomes to specify the location of origins of DNA replication. The S. cerevisiae ORC binds to specific DNA sequences throughout the cell ...The Origin Recognition Complex (ORC) binds to sites in chromosomes to specify the location of origins of DNA replication. The S. cerevisiae ORC binds to specific DNA sequences throughout the cell cycle but becomes active only when it binds to the replication initiator Cdc6. It has been unclear at the molecular level how Cdc6 activates ORC, converting it to an active recruiter of the Mcm2-7 hexamer, the core of the replicative helicase. Here we report the cryo-EM structure at 3.3 Å resolution of the yeast ORC-Cdc6 bound to an 85-bp ARS1 origin DNA. The structure reveals that Cdc6 contributes to origin DNA recognition via its winged helix domain (WHD) and its initiator-specific motif. Cdc6 binding rearranges a short α-helix in the Orc1 AAA+ domain and the Orc2 WHD, leading to the activation of the Cdc6 ATPase and the formation of the three sites for the recruitment of Mcm2-7, none of which are present in ORC alone. The results illuminate the molecular mechanism of a critical biochemical step in the licensing of eukaryotic replication origins.
History
DepositionApr 1, 2021-
Header (metadata) releaseJun 2, 2021-
Map releaseJun 2, 2021-
UpdateAug 4, 2021-
Current statusAug 4, 2021Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.014
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.014
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-7mca
  • Surface level: 0.014
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_23755.png

Downloads & links

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Map

FileDownload / File: emd_23755.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 0.828 Å
Density
Contour LevelBy AUTHOR: 0.011 / Movie #1: 0.014
Minimum - Maximum-0.06257838 - 0.09342496
Average (Standard dev.)0.00040741984 (±0.003358358)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions280280280
Spacing280280280
CellA=B=C: 231.84 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.8280.8280.828
M x/y/z280280280
origin x/y/z0.0000.0000.000
length x/y/z231.840231.840231.840
α/β/γ90.00090.00090.000
start NX/NY/NZ1219875
NX/NY/NZ141223232
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS280280280
D min/max/mean-0.0630.0930.000

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Supplemental data

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Half map: #2

Fileemd_23755_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_23755_half_map_2.map
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 origin replication complex with Cdc6 and ARS1 origin DNA

EntireName: Complex of origin replication complex with Cdc6 and ARS1 origin DNA
Components
  • Complex: Complex of origin replication complex with Cdc6 and ARS1 origin DNA
    • Protein or peptide: Origin recognition complex subunit 1
    • Protein or peptide: Origin recognition complex subunit 2
    • Protein or peptide: Origin recognition complex subunit 3
    • Protein or peptide: Origin recognition complex subunit 4
    • Protein or peptide: Origin recognition complex subunit 5
    • Protein or peptide: Origin recognition complex subunit 6
    • DNA: DNA (85-MER)
    • DNA: DNA (85-MER)
    • Protein or peptide: Cell division control protein 6
  • Ligand: MAGNESIUM ION
  • Ligand: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER

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Supramolecule #1: Complex of origin replication complex with Cdc6 and ARS1 origin DNA

SupramoleculeName: Complex of origin replication complex with Cdc6 and ARS1 origin DNA
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#9
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: unidentified baculovirus / Recombinant cell: Hi-5 cell / Recombinant plasmid: pAcUW51
Molecular weightTheoretical: 470 KDa

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Macromolecule #1: Origin recognition complex subunit 1

MacromoleculeName: Origin recognition complex subunit 1 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 104.546164 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MAKTLKDLQG WEIITTDEQG NIIDGGQKRL RRRGAKTEHY LKRSSDGIKL GRGDSVVMHN EAAGTYSVYM IQELRLNTLN NVVELWALT YLRWFEVNPL AHYRQFNPDA NILNRPLNYY NKLFSETANK NELYLTAELA ELQLFNFIRV ANVMDGSKWE V LKGNVDPE ...String:
MAKTLKDLQG WEIITTDEQG NIIDGGQKRL RRRGAKTEHY LKRSSDGIKL GRGDSVVMHN EAAGTYSVYM IQELRLNTLN NVVELWALT YLRWFEVNPL AHYRQFNPDA NILNRPLNYY NKLFSETANK NELYLTAELA ELQLFNFIRV ANVMDGSKWE V LKGNVDPE RDFTVRYICE PTGEKFVDIN IEDVKAYIKK VEPREAQEYL KDLTLPSKKK EIKRGPQKKD KATQTAQISD AE TRATDIT DNEDGNEDES SDYESPSDID VSEDMDSGEI SADELEEEED EEEDEDEEEK EARHTNSPRK RGRKIKLGKD DID ASVQPP PKKRGRKPKD PSKPRQMLLI SSCRANNTPV IRKFTKKNVA RAKKKYTPFS KRFKSIAAIP DLTSLPEFYG NSSE LMASR FENKLKTTQK HQIVETIFSK VKKQLNSSYV KEEILKSANF QDYLPARENE FASIYLSAYS AIESDSATTI YVAGT PGVG KTLTVREVVK ELLSSSAQRE IPDFLYVEIN GLKMVKPTDC YETLWNKVSG ERLTWAASME SLEFYFKRVP KNKKKT IVV LLDELDAMVT KSQDIMYNFF NWTTYENAKL IVIAVANTMD LPERQLGNKI TSRIGFTRIM FTGYTHEELK NIIDLRL KG LNDSFFYVDT KTGNAILIDA AGNDTTVKQT LPEDVRKVRL RMSADAIEIA SRKVASVSGD ARRALKVCKR AAEIAEKH Y MAKHGYGYDG KTVIEDENEE QIYDDEDKDL IESNKAKDDN DDDDDNDGVQ TVHITHVMKA LNETLNSHVI TFMTRLSFT AKLFIYALLN LMKKNGSQEQ ELGDIVDEIK LLIEVNGSNK FVMEIAKTLF QQGSDNISEQ LRIISWDFVL NQLLDAGILF KQTMKNDRI CCVKLNISVE EAKRAMNEDE TLRNL

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Macromolecule #2: Origin recognition complex subunit 2

MacromoleculeName: Origin recognition complex subunit 2 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 71.34218 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MLNGEDFVEH NDILSSPAKS RNVTPKRVDP HGERQLRRIH SSKKNLLERI SLVGNERKNT SPDPALKPKT PSKAPRKRGR PRKIQEELT DRIKKDEKDT ISSKKKRKLD KDTSGNVNEE SKTSNNKQVM EKTGIKEKRE REKIQVATTT YEDNVTPQTD D NFVSNSPE ...String:
MLNGEDFVEH NDILSSPAKS RNVTPKRVDP HGERQLRRIH SSKKNLLERI SLVGNERKNT SPDPALKPKT PSKAPRKRGR PRKIQEELT DRIKKDEKDT ISSKKKRKLD KDTSGNVNEE SKTSNNKQVM EKTGIKEKRE REKIQVATTT YEDNVTPQTD D NFVSNSPE PPEPATPSKK SLTTNHDFTS PLKQIIMNNL KEYKDSTSPG KLTLSRNFTP TPVPKNKKLY QTSETKSASS FL DTFEGYF DQRKIVRTNA KSRHTMSMAP DVTREEFSLV SNFFNENFQK RPRQKLFEIQ KKMFPQYWFE LTQGFSLLFY GVG SKRNFL EEFAIDYLSP KIAYSQLAYE NELQQNKPVN SIPCLILNGY NPSCNYRDVF KEITDLLVPA ELTRSETKYW GNHV ILQIQ KMIDFYKNQP LDIKLILVVH NLDGPSIRKN TFQTMLSFLS VIRQIAIVAS TDHIYAPLLW DNMKAQNYNF VFHDI SNFE PSTVESTFQD VMKMGKSDTS SGAEGAKYVL QSLTVNSKKM YKLLIETQMQ NMGNLSANTG PKRGTQRTGV ELKLFN HLC AADFIASNEI ALRSMLREFI EHKMANITKN NSGMEIIWVP YTYAELEKLL KTVLNTL

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Macromolecule #3: Origin recognition complex subunit 3

MacromoleculeName: Origin recognition complex subunit 3 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 72.161766 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MSDLNQSKKM NVSEFADAQR SHYTVYPSLP QSNKNDKHIP FVKLLSGKES EVNVEKRWEL YHQLHSHFHD QVDHIIDNIE ADLKAEISD LLYSETTQKR RCFNTIFLLG SDSTTKIELK DESSRYNVLI ELTPKESPNV RMMLRRSMYK LYSAADAEEH P TIKYEDIN ...String:
MSDLNQSKKM NVSEFADAQR SHYTVYPSLP QSNKNDKHIP FVKLLSGKES EVNVEKRWEL YHQLHSHFHD QVDHIIDNIE ADLKAEISD LLYSETTQKR RCFNTIFLLG SDSTTKIELK DESSRYNVLI ELTPKESPNV RMMLRRSMYK LYSAADAEEH P TIKYEDIN DEDGDFTEQN NDVSYDLSLV ENFKRLFGKD LAMVFNFKDV DSINFNTLDN FIILLKSAFK YDHVKISLIF NI NTNLSNI EKNLRQSTIR LLKRNYHKLD VSSNKGFKYG NQIFQSFLDT VDGKLNLSDR FVEFILSKMA NNTNHNLQLL TKM LDYSLM SYFFQNAFSV FIDPVNVDFL NDDYLKILSR CPTFMFFVEG LIKQHAPADE ILSLLTNKNR GLEEFFVEFL VREN PINGH AKFVARFLEE ELNITNFNLI ELYHNLLIGK LDSYLDRWSA CKEYKDRLHF EPIDTIFQEL FTLDNRSGLL TQSIF PSYK SNIEDNLLSW EQVLPSLDKE NYDTLSGDLD KIMAPVLGQL FKLYREANMT INIYDFYIAF RETLPKEEIL NFIRKD PSN TKLLELAETP DAFDKVALIL FMQAIFAFEN MGLIKFQSTK SYDLVEKCVW RGI

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Macromolecule #4: Origin recognition complex subunit 4

MacromoleculeName: Origin recognition complex subunit 4 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 60.772152 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MTISEARLSP QVNLLPIKRH SNEEVEETAA ILKKRTIDNE KCKDSDPGFG SLQRRLLQQL YGTLPTDEKI IFTYLQDCQQ EIDRIIKQS IIQKESHSVI LVGPRQSYKT YLLDYELSLL QQSYKEQFIT IRLNGFIHSE QTAINGIATQ LEQQLQKIHG S EEKIDDTS ...String:
MTISEARLSP QVNLLPIKRH SNEEVEETAA ILKKRTIDNE KCKDSDPGFG SLQRRLLQQL YGTLPTDEKI IFTYLQDCQQ EIDRIIKQS IIQKESHSVI LVGPRQSYKT YLLDYELSLL QQSYKEQFIT IRLNGFIHSE QTAINGIATQ LEQQLQKIHG S EEKIDDTS LETISSGSLT EVFEKILLLL DSTTKTRNED SGEVDRESIT KITVVFIFDE IDTFAGPVRQ TLLYNLFDMV EH SRVPVCI FGCTTKLNIL EYLEKRVKSR FSQRVIYMPQ IQNLDDMVDA VRNLLTVRSE ISPWVSQWNE TLEKELSDPR SNL NRHIRM NFETFRSLPT LKNSIIPLVA TSKNFGSLCT AIKSCSFLDI YNKNQLSNNL TGRLQSLSDL ELAILISAAR VALR AKDGS FNFNLAYAEY EKMIKAINSR IPTVAPTTNV GTGQSTFSID NTIKLWLKKD VKNVWENLVQ LDFFTEKSAV GLRDN ATAA FYASNYQFQG TMIPFDLRSY QMQIILQELR RIIPKSNMYY SWTQL

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Macromolecule #5: Origin recognition complex subunit 5

MacromoleculeName: Origin recognition complex subunit 5 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 55.347168 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MNVTTPEVAF REYQTNCLAS YISADPDITP SNLILQGYSG TGKTYTLKKY FNANPNLHAV WLEPVELVSW KPLLQAIART VQYKLKTLY PNIPTTDYDP LQVEEPFLLV KTLHNIFVQY ESLQEKTCLF LILDGFDSLQ DLDAALFNKY IKLNELLPKD S KINIKFIY ...String:
MNVTTPEVAF REYQTNCLAS YISADPDITP SNLILQGYSG TGKTYTLKKY FNANPNLHAV WLEPVELVSW KPLLQAIART VQYKLKTLY PNIPTTDYDP LQVEEPFLLV KTLHNIFVQY ESLQEKTCLF LILDGFDSLQ DLDAALFNKY IKLNELLPKD S KINIKFIY TMLETSFLQR YSTHCIPTVM FPRYNVDEVS TILVMSRCGE LMEDSCLRKR IIEEQITDCT DDQFQNVAAN FI HLIVQAF HSYTGNDIFA LNDLIDFKWP KYVSRITKEN IFEPLALYKS AIKLFLSTDD NLSENGQGES AITTNRDDLE NSQ TYDLSI ISKYLLIASY ICSYLEPRYD ASIFSRKTRI IQGRAAYGRR KKKEVNPRYL QPSLFAIERL LAIFQAIFPI QGKA ESGSL SALREESLMK ANIEVFQNLS ELHTLKLIAT TMNKNIDYLS PKVRWKVNVP WEIIKEISES VHFNISDYFS DIHE

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Macromolecule #6: Origin recognition complex subunit 6

MacromoleculeName: Origin recognition complex subunit 6 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 50.369531 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MSMQQVQHCV AEVLRLDPQE KPDWSSGYLK KLTNATSILY NTSLNKVMLK QDEEVARCHI CAYIASQKMN EKHMPDLCYY IDSIPLEPK KAKHLMNLFR QSLSNSSPMK QFAWTPSPKK NKRSPVKNGG RFTSSDPKEL RNQLFGTPTK VRKSQNNDSF V IPELPPMQ ...String:
MSMQQVQHCV AEVLRLDPQE KPDWSSGYLK KLTNATSILY NTSLNKVMLK QDEEVARCHI CAYIASQKMN EKHMPDLCYY IDSIPLEPK KAKHLMNLFR QSLSNSSPMK QFAWTPSPKK NKRSPVKNGG RFTSSDPKEL RNQLFGTPTK VRKSQNNDSF V IPELPPMQ TNESPSITRR KLAFEEDEDE DEEEPGNDGL SLKSHSNKSI TGTRNVDSDE YENHESDPTS EEEPLGVQES RS GRTKQNK AVGKPQSELK TAKALRKRGR IPNSLLVKKY CKMTTEEIIR LCNDFELPRE VAYKIVDEYN INASRLVCPW QLV CGLVLN CTFIVFNERR RKDPRIDHFI VSKMCSLMLT SKVDDVIECV KLVKELIIGE KWFRDLQIRY DDFDGIRYDE IIFR KLGSM LQTTNILVTD DQYNIWKKRI EMDLALTEPL

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Macromolecule #9: Cell division control protein 6

MacromoleculeName: Cell division control protein 6 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 58.112367 KDa
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MSAIPITPTK RIRRNLFDDA PATPPRPLKR KKLQFTDVTP ESSPEKLQFG SQSIFLRTKA LLQKSSELVN LNSSDGALPA RTAEYEQVM NFLAKAISEH RSDSLYITGP PGTGKTAQLD MIIRQKFQSL PLSLSTPRSK DVLRHTNPNL QNLSWFELPD G RLESVAVT ...String:
MSAIPITPTK RIRRNLFDDA PATPPRPLKR KKLQFTDVTP ESSPEKLQFG SQSIFLRTKA LLQKSSELVN LNSSDGALPA RTAEYEQVM NFLAKAISEH RSDSLYITGP PGTGKTAQLD MIIRQKFQSL PLSLSTPRSK DVLRHTNPNL QNLSWFELPD G RLESVAVT SINCISLGEP SSIFQKIFDS FQDLNGPTLQ IKNMQHLQKF LEPYHKKTTF VVVLDEMDRL LHANTSETQS VR TILELFL LAKLPTVSFV LIGMANSLDM KDRFLSRLNL DRGLLPQTIV FQPYTAEQMY EIVIQKMSSL PTIIFQPMAI KFA AKKCAG NTGDLRKLFD VLRGSIEIYE LEKRFLLSPT RGSLNSAQVP LTPTTSPVKK SYPEPQGKIG LNYIAKVFSK FVNN NSTRT RIAKLNIQQK LILCTIIQSL KLNSDATIDE SFDHYIKAIT KTDTLAPLQR NEFLEICTIL ETCGLVSIKK TKCKG KTKR FVDKIDVDLD MREFYDEMTK ISILKPFLH

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Macromolecule #7: DNA (85-MER)

MacromoleculeName: DNA (85-MER) / type: dna / ID: 7 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 26.184838 KDa
SequenceString: (DG)(DT)(DT)(DA)(DT)(DT)(DT)(DT)(DA)(DC) (DA)(DG)(DA)(DT)(DT)(DT)(DT)(DA)(DT)(DG) (DT)(DT)(DT)(DA)(DG)(DA)(DT)(DC)(DT) (DT)(DT)(DT)(DA)(DT)(DG)(DC)(DT)(DT)(DG) (DC) (DT)(DT)(DT)(DT)(DC)(DA) ...String:
(DG)(DT)(DT)(DA)(DT)(DT)(DT)(DT)(DA)(DC) (DA)(DG)(DA)(DT)(DT)(DT)(DT)(DA)(DT)(DG) (DT)(DT)(DT)(DA)(DG)(DA)(DT)(DC)(DT) (DT)(DT)(DT)(DA)(DT)(DG)(DC)(DT)(DT)(DG) (DC) (DT)(DT)(DT)(DT)(DC)(DA)(DA)(DA) (DA)(DG)(DG)(DC)(DC)(DT)(DG)(DC)(DA)(DG) (DG)(DC) (DA)(DA)(DG)(DT)(DG)(DC)(DA) (DC)(DA)(DA)(DA)(DC)(DA)(DA)(DT)(DA)(DC) (DT)(DT)(DA) (DA)(DA)(DT)(DA)(DA)

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Macromolecule #8: DNA (85-MER)

MacromoleculeName: DNA (85-MER) / type: dna / ID: 8 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 26.229906 KDa
SequenceString: (DT)(DT)(DA)(DT)(DT)(DT)(DA)(DA)(DG)(DT) (DA)(DT)(DT)(DG)(DT)(DT)(DT)(DG)(DT)(DG) (DC)(DA)(DC)(DT)(DT)(DG)(DC)(DC)(DT) (DG)(DC)(DA)(DG)(DG)(DC)(DC)(DT)(DT)(DT) (DT) (DG)(DA)(DA)(DA)(DA)(DG) ...String:
(DT)(DT)(DA)(DT)(DT)(DT)(DA)(DA)(DG)(DT) (DA)(DT)(DT)(DG)(DT)(DT)(DT)(DG)(DT)(DG) (DC)(DA)(DC)(DT)(DT)(DG)(DC)(DC)(DT) (DG)(DC)(DA)(DG)(DG)(DC)(DC)(DT)(DT)(DT) (DT) (DG)(DA)(DA)(DA)(DA)(DG)(DC)(DA) (DA)(DG)(DC)(DA)(DT)(DA)(DA)(DA)(DA)(DG) (DA)(DT) (DC)(DT)(DA)(DA)(DA)(DC)(DA) (DT)(DA)(DA)(DA)(DA)(DT)(DC)(DT)(DG)(DT) (DA)(DA)(DA) (DA)(DT)(DA)(DA)(DC)

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Macromolecule #10: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 10 / Number of copies: 4 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #11: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER

MacromoleculeName: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / type: ligand / ID: 11 / Number of copies: 4 / Formula: AGS
Molecular weightTheoretical: 523.247 Da
Chemical component information

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

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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
Component:
ConcentrationFormulaName
50.0 mMHEPES
100.0 mMCH3CO2Kpotassium acetate
5.0 mMC4H6MgO4magnesium acetate
5.0 mMATP-gamma-S
2.0 mMDTT
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE
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: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 76.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 177397
CTF correctionSoftware - Name: CTFFIND (ver. 4.1.10)
Startup modelType of model: INSILICO MODEL / In silico model: cryosparc ab initio reconstruction
Initial angle assignmentType: OTHER / Software - Name: cryoSPARC (ver. 2)
Final 3D classificationNumber classes: 2 / Avg.num./class: 1 / Software - Name: RELION (ver. 3.0)
Final angle assignmentType: OTHER / Software - Name: cryoSPARC (ver. 2)
Final reconstructionNumber classes used: 1 / Resolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 72000
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

5v8f

RefinementSpace: REAL / Protocol: OTHER / Overall B value: 118.5 / Target criteria: correlation coefficient
Output model

PDB-7mca:
Structure of the S. cerevisiae origin recognition complex bound to the replication initiator Cdc6 and the ARS1 origin DNA.

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Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

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