[English] 日本語
Yorodumi- EMDB-21701: Structure of the Saccharomyces cerevisiae polymerase epsilon holo... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21701 | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | Structure of the Saccharomyces cerevisiae polymerase epsilon holoenzyme | ||||||||||||
Map data | Saccharomyces cerevisiae polymerase epsilon holoenzyme | ||||||||||||
Sample |
| ||||||||||||
Function / homology | Function and homology information : / : / chromatin remodeling => GO:0006338 / CHRAC / : / DNA-templated DNA replication maintenance of fidelity / gene conversion / DNA replication initiation / epsilon DNA polymerase complex / SUMO binding ...: / : / chromatin remodeling => GO:0006338 / CHRAC / : / DNA-templated DNA replication maintenance of fidelity / gene conversion / DNA replication initiation / epsilon DNA polymerase complex / SUMO binding / Activation of the pre-replicative complex / single-stranded DNA 3'-5' DNA exonuclease activity / Termination of translesion DNA synthesis / mitotic DNA replication checkpoint signaling / mitotic intra-S DNA damage checkpoint signaling / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / nucleotide-excision repair, DNA gap filling / DNA replication proofreading / mitotic sister chromatid cohesion / leading strand elongation / nuclear replication fork / Dual incision in TC-NER / error-prone translesion synthesis / heterochromatin formation / nucleosomal DNA binding / base-excision repair, gap-filling / replication fork / base-excision repair / chromatin DNA binding / DNA-templated DNA replication / double-strand break repair via nonhomologous end joining / double-strand break repair / mitotic cell cycle / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / double-stranded DNA binding / DNA-directed DNA polymerase / DNA-directed DNA polymerase activity / cell cycle / protein heterodimerization activity / nucleotide binding / mRNA binding / DNA damage response / DNA binding / zinc ion binding / nucleus / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.5 Å | ||||||||||||
Authors | Yuan Z / Georgescu R / Schauer GD / O'Donnell M / Li H | ||||||||||||
Funding support | United States, 3 items
| ||||||||||||
Citation | Journal: Nat Commun / Year: 2020 Title: Structure of the polymerase ε holoenzyme and atomic model of the leading strand replisome. Authors: Zuanning Yuan / Roxana Georgescu / Grant D Schauer / Michael E O'Donnell / Huilin Li / Abstract: The eukaryotic leading strand DNA polymerase (Pol) ε contains 4 subunits, Pol2, Dpb2, Dpb3 and Dpb4. Pol2 is a fusion of two B-family Pols; the N-terminal Pol module is catalytic and the C-terminal ...The eukaryotic leading strand DNA polymerase (Pol) ε contains 4 subunits, Pol2, Dpb2, Dpb3 and Dpb4. Pol2 is a fusion of two B-family Pols; the N-terminal Pol module is catalytic and the C-terminal Pol module is non-catalytic. Despite extensive efforts, there is no atomic structure for Pol ε holoenzyme, critical to understanding how DNA synthesis is coordinated with unwinding and the DNA path through the CMG helicase-Pol ε-PCNA clamp. We show here a 3.5-Å cryo-EM structure of yeast Pol ε revealing that the Dpb3-Dpb4 subunits bridge the two DNA Pol modules of Pol2, holding them rigid. This information enabled an atomic model of the leading strand replisome. Interestingly, the model suggests that an OB fold in Dbp2 directs leading ssDNA from CMG to the Pol ε active site. These results complete the DNA path from entry of parental DNA into CMG to exit of daughter DNA from PCNA. | ||||||||||||
History |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_21701.map.gz | 6.6 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-21701-v30.xml emd-21701.xml | 16.9 KB 16.9 KB | Display Display | EMDB header |
Images | emd_21701.png | 144.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21701 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21701 | HTTPS FTP |
-Validation report
Summary document | emd_21701_validation.pdf.gz | 345.7 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_21701_full_validation.pdf.gz | 345.3 KB | Display | |
Data in XML | emd_21701_validation.xml.gz | 6.1 KB | Display | |
Data in CIF | emd_21701_validation.cif.gz | 6.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21701 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21701 | HTTPS FTP |
-Related structure data
Related structure data | 6wjvMC M: atomic model generated by this map C: citing same article (ref.) |
---|---|
Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_21701.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Saccharomyces cerevisiae polymerase epsilon holoenzyme | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.029 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
-Entire : DNA polymerase epsilon
Entire | Name: DNA polymerase epsilon |
---|---|
Components |
|
-Supramolecule #1: DNA polymerase epsilon
Supramolecule | Name: DNA polymerase epsilon / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4 |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Strain: ATCC 204508 / S288c |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) / Recombinant plasmid: pRS |
-Macromolecule #1: DNA polymerase epsilon catalytic subunit A
Macromolecule | Name: DNA polymerase epsilon catalytic subunit A / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed DNA polymerase |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Strain: ATCC 204508 / S288c |
Molecular weight | Theoretical: 255.992484 KDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Sequence | String: MMFGKKKNNG GSSTARYSAG NKYNTLSNNY ALSAQQLLNA SKIDDIDSMM GFERYVPPQY NGRFDAKDID QIPGRVGWLT NMHATLVSQ ETLSSGSNGG GNSNDGERVT TNQGISGVDF YFLDEEGGSF KSTVVYDPYF FIACNDESRV NDVEELVKKY L ESCLKSLQ ...String: MMFGKKKNNG GSSTARYSAG NKYNTLSNNY ALSAQQLLNA SKIDDIDSMM GFERYVPPQY NGRFDAKDID QIPGRVGWLT NMHATLVSQ ETLSSGSNGG GNSNDGERVT TNQGISGVDF YFLDEEGGSF KSTVVYDPYF FIACNDESRV NDVEELVKKY L ESCLKSLQ IIRKEDLTMD NHLLGLQKTL IKLSFVNSNQ LFEARKLLRP ILQDNANNNV QRNIYNVAAN GSEKVDAKHL IE DIREYDV PYHVRVSIDK DIRVGKWYKV TQQGFIEDTR KIAFADPVVM AFDIETTKPP LKFPDSAVDQ IMMISYMIDG EGF LITNRE IISEDIEDFE YTPKPEYPGF FTIFNENDEV ALLQRFFEHI RDVRPTVIST FNGDFFDWPF IHNRSKIHGL DMFD EIGFA PDAEGEYKSS YCSHMDCFRW VKRDSYLPQG SQGLKAVTQS KLGYNPIELD PELMTPYAFE KPQHLSEYSV SDAVA TYYL YMKYVHPFIF SLCTIIPLNP DETLRKGTGT LCEMLLMVQA YQHNILLPNK HTDPIERFYD GHLLESETYV GGHVES LEA GVFRSDLKNE FKIDPSAIDE LLQELPEALK FSVEVENKSS VDKVTNFEEI KNQITQKLLE LKENNIRNEL PLIYHVD VA SMYPNIMTTN RLQPDSIKAE RDCASCDFNR PGKTCARKLK WAWRGEFFPS KMDEYNMIKR ALQNETFPNK NKFSKKKV L TFDELSYADQ VIHIKKRLTE YSRKVYHRVK VSEIVEREAI VCQRENPFYV DTVKSFRDRR YEFKGLAKTW KGNLSKIDP SDKHARDEAK KMIVLYDSLQ LAHKVILNSF YGYVMRKGSR WYSMEMAGIT CLTGATIIQM ARALVERVGR PLELDTDGIW CILPKSFPE TYFFTLENGK KLYLSYPCSM LNYRVHQKFT NHQYQELKDP LNYIYETHSE NTIFFEVDGP YKAMILPSSK E EGKGIKKR YAVFNEDGSL AELKGFELKR RGELQLIKNF QSDIFKVFLE GDTLEGCYSA VASVCNRWLD VLDSHGLMLE DE DLVSLIC ENRSMSKTLK EYEGQKSTSI TTARRLGDFL GEDMVKDKGL QCKYIISSKP FNAPVTERAI PVAIFSADIP IKR SFLRRW TLDPSLEDLD IRTIIDWGYY RERLGSAIQK IITIPAALQG VSNPVPRVEH PDWLKRKIAT KEDKFKQTSL TKFF SKTKN VPTMGKIKDI EDLFEPTVEE DNAKIKIART TKKKAVSKRK RNQLTNEEDP LVLPSEIPSM DEDYVGWLNY QKIKW KIQA RDRKRRDQLF GNTNSSRERS ALGSMIRKQA ESYANSTWEV LQYKDSGEPG VLEVFVTING KVQNITFHIP KTIYMK FKS QTMPLQKIKN CLIEKSSASL PNNPKTSNPA GGQLFKITLP ESVFLEEKEN CTSIFNDENV LGVFEGTITP HQRAIMD LG ASVTFRSKAM GALGKGIQQG FEMKDLSMAE NERYLSGFSM DIGYLLHFPT SIGYEFFSLF KSWGDTITIL VLKPSNQA Q EINASSLGQI YKQMFEKKKG KIETYSYLVD IKEDINFEFV YFTDISKLYR RLSQETTKLK EERGLQFLLL LQSPFITKL LGTIRLLNQM PIVKLSLNEV LLPQLNWQPT LLKKLVNHVL SSGSWISHLI KLSQYSNIPI CNLRLDSMDY IIDVLYARKL KKENIVLWW NEKAPLPDHG GIQNDFDLNT SWIMNDSEFP KINNSGVYDN VVLDVGVDNL TVNTILTSAL INDAEGSDLV N NNMGIDDK DAVINSPSEF VHDAFSNDAL NVLRGMLKEW WDEALKENST ADLLVNSLAS WVQNPNAKLF DGLLRYHVHN LT KKALLQL VNEFSALGST IVYADRNQIL IKTNKYSPEN CYAYSQYMMK AVRTNPMFSY LDLNIKRYWD LLIWMDKFNF SGL ACIEIE EKENQDYTAV SQWQLKKFLS PIYQPEFEDW MMIILDSMLK TKQSYLKLNS GTQRPTQIVN VKKQDKEDSV ENSL NGFSH LFSKPLMKRV KKLFKNQQEF ILDPQYEADY VIPVLPGSHL NVKNPLLELV KSLCHVMLLS KSTILEIRTL RKELL KIFE LREFAKVAEF KDPSLSLVVP DFLCEYCFFI SDIDFCKAAP ESIFSCVRCH KAFNQVLLQE HLIQKLRSDI ESYLIQ DLR CSRCHKVKRD YMSAHCPCAG AWEGTLPRES IVQKLNVFKQ VAKYYGFDIL LSCIADLTI |
-Macromolecule #2: DNA polymerase epsilon subunit B
Macromolecule | Name: DNA polymerase epsilon subunit B / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Strain: ATCC 204508 / S288c |
Molecular weight | Theoretical: 78.425852 KDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Sequence | String: MFGSGNVLPV KIQPPLLRPL AYRVLSRKYG LSIKSDGLSA LAEFVGTNIG ANWRQGPATI KFLEQFAAVW KQQERGLFID QSGVKEVIQ EMKEREKVEW SHEHPIQHEE NILGRTDDDE NNSDDEMPIA ADSSLQNVSL SSPMRQPTER DEYKQPFKPE S SKALDWRD ...String: MFGSGNVLPV KIQPPLLRPL AYRVLSRKYG LSIKSDGLSA LAEFVGTNIG ANWRQGPATI KFLEQFAAVW KQQERGLFID QSGVKEVIQ EMKEREKVEW SHEHPIQHEE NILGRTDDDE NNSDDEMPIA ADSSLQNVSL SSPMRQPTER DEYKQPFKPE S SKALDWRD YFKVINASQQ QRFSYNPHKM QFIFVPNKKQ NGLGGIAGFL PDIEDKVQMF LTRYYLTNDR VMRNENFQNS DM FNPLSSM VSLQNELSNT NRQQQSSSMS ITPIKNLLGR DAQNFLLLGL LNKNFKGNWS LEDPSGSVEI DISQTIPTQG HYY VPGCMV LVEGIYYSVG NKFHVTSMTL PPGERREITL ETIGNLDLLG IHGISNNNFI ARLDKDLKIR LHLLEKELTD HKFV ILGAN LFLDDLKIMT ALSKILQKLN DDPPTLLIWQ GSFTSVPVFA SMSSRNISSS TQFKNNFDAL ATLLSRFDNL TENTT MIFI PGPNDLWGSM VSLGASGTLP QDPIPSAFTK KINKVCKNVV WSSNPTRIAY LSQEIVIFRD DLSGRFKRHR LEFPFN ESE DVYTENDNMM SKDTDIVPID ELVKEPDQLP QKVQETRKLV KTILDQGHLS PFLDSLRPIS WDLDHTLTLC PIPSTMV LC DTTSAQFDLT YNGCKVINPG SFIHNRRARY MEYVPSSKKT IQEEIYI |
-Macromolecule #3: DNA polymerase epsilon subunit C
Macromolecule | Name: DNA polymerase epsilon subunit C / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Strain: ATCC 204508 / S288c |
Molecular weight | Theoretical: 22.694014 KDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Sequence | String: MSNLVKEKAP VFPISKVKKI AKCDPEYVIT SNVAISATAF AAELFVQNLV EESLVLAQLN SKGKTSLRLS LNSIEECVEK RDNFRFLED AIKQLKKNSA LDKKRELNMQ PGRSDQEVVI EEPELHEDDG VEEEEEEDEV SEEEEPVHNE ELLDDSKDQQ N DKSTRSVA ...String: MSNLVKEKAP VFPISKVKKI AKCDPEYVIT SNVAISATAF AAELFVQNLV EESLVLAQLN SKGKTSLRLS LNSIEECVEK RDNFRFLED AIKQLKKNSA LDKKRELNMQ PGRSDQEVVI EEPELHEDDG VEEEEEEDEV SEEEEPVHNE ELLDDSKDQQ N DKSTRSVA SLLSRFQYKS ALDVGEHSDS SDIEVDHTKS TDP |
-Macromolecule #4: DNA polymerase epsilon subunit D
Macromolecule | Name: DNA polymerase epsilon subunit D / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) Strain: ATCC 204508 / S288c |
Molecular weight | Theoretical: 22.029484 KDa |
Recombinant expression | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Sequence | String: MPPKGWRKDA QGNYPTTSYI KEQENITIQD LLFPKSTIVN LAREVPQQSG KKLLINKDAS LALQRGATVF VNHLLLFARE IAKSQDKKS CSVDDVLSAL DHIGHSALKG PVRDKLDEYQ AAVEQRKKEK LDSGEVDADG DIDMGEDKEN VPVEKVKEHD E IEEQGDAL ...String: MPPKGWRKDA QGNYPTTSYI KEQENITIQD LLFPKSTIVN LAREVPQQSG KKLLINKDAS LALQRGATVF VNHLLLFARE IAKSQDKKS CSVDDVLSAL DHIGHSALKG PVRDKLDEYQ AAVEQRKKEK LDSGEVDADG DIDMGEDKEN VPVEKVKEHD E IEEQGDAL QDVEESSEKK QKTESQDVET RVQNLEQT |
-Macromolecule #5: ZINC ION
Macromolecule | Name: ZINC ION / type: ligand / ID: 5 / Number of copies: 1 / Formula: ZN |
---|---|
Molecular weight | Theoretical: 65.409 Da |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
---|---|
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 60.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.5 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 187298 |
---|---|
Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: FLEXIBLE FIT |
---|---|
Output model | PDB-6wjv: |