[English] 日本語
Yorodumi
- EMDB-29862: Partial auto-inhibitory complex of Xenopus laevis DNA polymerase ... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: EMDB / ID: EMD-29862
TitlePartial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase
Map dataPartial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase
Sample
  • Complex: Polymerase alpha-primase
    • Complex: Polymerase alpha
      • Protein or peptide: DNA polymerase alpha catalytic subunit
      • Protein or peptide: DNA polymerase alpha subunit B
    • Complex: Primase
      • Protein or peptide: DNA primase large subunit
  • Ligand: ZINC ION
  • Ligand: IRON/SULFUR CLUSTER
KeywordsPrimase / DNA polymerase / chimeric RNA-DNA primer / RNA/DNA hybrid / DNA replication / DNA synthesis / REPLICATION / TRANSFERASE
Function / homology
Function and homology information


alpha DNA polymerase:primase complex / primosome complex / DNA replication, synthesis of primer / lagging strand elongation / mitotic DNA replication initiation / DNA strand elongation involved in DNA replication / leading strand elongation / DNA replication initiation / double-strand break repair via nonhomologous end joining / nuclear matrix ...alpha DNA polymerase:primase complex / primosome complex / DNA replication, synthesis of primer / lagging strand elongation / mitotic DNA replication initiation / DNA strand elongation involved in DNA replication / leading strand elongation / DNA replication initiation / double-strand break repair via nonhomologous end joining / nuclear matrix / nuclear envelope / 4 iron, 4 sulfur cluster binding / DNA replication / DNA-directed DNA polymerase / DNA-directed DNA polymerase activity / nucleotide binding / chromatin binding / chromatin / nucleolus / DNA binding / nucleoplasm / nucleus / metal ion binding
Similarity search - Function
DNA polymerase alpha, subunit B, N-terminal domain superfamily / : / DNA polymerase alpha subunit B, OB domain / DNA polymerase alpha subunit B N-terminal / DNA polymerase alpha, subunit B, N-terminal / DNA polymerase alpha, subunit B / DNA primase, large subunit, eukaryotic / DNA primase large subunit, eukaryotic/archaeal / DNA polymerase alpha catalytic subunit, N-terminal domain / DNA polymerase alpha, zinc finger domain superfamily ...DNA polymerase alpha, subunit B, N-terminal domain superfamily / : / DNA polymerase alpha subunit B, OB domain / DNA polymerase alpha subunit B N-terminal / DNA polymerase alpha, subunit B, N-terminal / DNA polymerase alpha, subunit B / DNA primase, large subunit, eukaryotic / DNA primase large subunit, eukaryotic/archaeal / DNA polymerase alpha catalytic subunit, N-terminal domain / DNA polymerase alpha, zinc finger domain superfamily / Eukaryotic and archaeal DNA primase, large subunit / DNA Polymerase alpha zinc finger / DNA polymerase alpha subunit p180 N terminal / Zinc finger, DNA-directed DNA polymerase, family B, alpha / DNA polymerase alpha catalytic subunit, catalytic domain / DNA polymerase alpha/delta/epsilon, subunit B / DNA polymerase alpha/epsilon subunit B / DNA polymerase family B, thumb domain / DNA-directed DNA polymerase, family B, multifunctional domain / DNA polymerase family B signature. / DNA-directed DNA polymerase, family B, conserved site / DNA polymerase family B / DNA polymerase family B, exonuclease domain / DNA-directed DNA polymerase, family B, exonuclease domain / DNA polymerase, palm domain superfamily / DNA polymerase type-B family / DNA-directed DNA polymerase, family B / Ribonuclease H superfamily / Ribonuclease H-like superfamily / DNA/RNA polymerase superfamily
Similarity search - Domain/homology
DNA primase large subunit / DNA polymerase alpha subunit B / DNA polymerase alpha catalytic subunit
Similarity search - Component
Biological speciesXenopus laevis (African clawed frog)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.8 Å
AuthorsMullins EA / Chazin WJ / Eichman BF
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM136401 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM118089 United States
National Institutes of Health/National Cancer Institute (NIH/NCI)PO1CA92584 United States
CitationJournal: bioRxiv / Year: 2023
Title: A mechanistic model of primer synthesis from catalytic structures of DNA polymerase α-primase.
Authors: Elwood A Mullins / Lauren E Salay / Clarissa L Durie / Noah P Bradley / Jane E Jackman / Melanie D Ohi / Walter J Chazin / Brandt F Eichman
Abstract: The mechanism by which polymerase α-primase (polα-primase) synthesizes chimeric RNA-DNA primers of defined length and composition, necessary for replication fidelity and genome stability, is ...The mechanism by which polymerase α-primase (polα-primase) synthesizes chimeric RNA-DNA primers of defined length and composition, necessary for replication fidelity and genome stability, is unknown. Here, we report cryo-EM structures of polα-primase in complex with primed templates representing various stages of DNA synthesis. Our data show how interaction of the primase regulatory subunit with the primer 5'-end facilitates handoff of the primer to polα and increases polα processivity, thereby regulating both RNA and DNA composition. The structures detail how flexibility within the heterotetramer enables synthesis across two active sites and provide evidence that termination of DNA synthesis is facilitated by reduction of polα and primase affinities for the varied conformations along the chimeric primer/template duplex. Together, these findings elucidate a critical catalytic step in replication initiation and provide a comprehensive model for primer synthesis by polα-primase.
History
DepositionFeb 21, 2023-
Header (metadata) releaseApr 12, 2023-
Map releaseApr 12, 2023-
UpdateMay 29, 2024-
Current statusMay 29, 2024Processing site: RCSB / Status: Released

-
Structure visualization

Supplemental images

emd_29862.png

Downloads & links

-
Map

FileDownload / File: emd_29862.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationPartial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.82 Å/pix.
x 400 pix.
= 328. Å		0.82 Å/pix.
x 400 pix.
= 328. Å		0.82 Å/pix.
x 400 pix.
= 328. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.82 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.01
Minimum - Maximum-0.0152480425 - 0.090278186
Average (Standard dev.)0.00004684871 (±0.0014017728)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 328.0 Å
α=β=γ: 90.0 °

-
Supplemental data

-
Additional map: Partial auto-inhibitory complex of Xenopus laevis DNA polymerase...

Fileemd_29862_additional_1.map
AnnotationPartial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase (locally sharpened)
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Half map: Partial auto-inhibitory complex of Xenopus laevis DNA polymerase...

Fileemd_29862_half_map_1.map
AnnotationPartial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase (half 1)
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Half map: Partial auto-inhibitory complex of Xenopus laevis DNA polymerase...

Fileemd_29862_half_map_2.map
AnnotationPartial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase (half 2)
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Sample components

-
Entire : Polymerase alpha-primase

EntireName: Polymerase alpha-primase
Components
  • Complex: Polymerase alpha-primase
    • Complex: Polymerase alpha
      • Protein or peptide: DNA polymerase alpha catalytic subunit
      • Protein or peptide: DNA polymerase alpha subunit B
    • Complex: Primase
      • Protein or peptide: DNA primase large subunit
  • Ligand: ZINC ION
  • Ligand: IRON/SULFUR CLUSTER

-
Supramolecule #1: Polymerase alpha-primase

SupramoleculeName: Polymerase alpha-primase / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 / Details: Heterotetrameric protein complex
Molecular weightTheoretical: 110 KDa

-
Supramolecule #2: Polymerase alpha

SupramoleculeName: Polymerase alpha / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#2 / Details: Heterodimer consisting of POLA1 and POLA2
Source (natural)Organism: Xenopus laevis (African clawed frog)
Molecular weightTheoretical: 200 KDa

-
Supramolecule #3: Primase

SupramoleculeName: Primase / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #3 / Details: Heterodimer consisting of PRIM1 and PRIM2
Source (natural)Organism: Xenopus laevis (African clawed frog)

-
Macromolecule #1: DNA polymerase alpha catalytic subunit

MacromoleculeName: DNA polymerase alpha catalytic subunit / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed DNA polymerase
Source (natural)Organism: Xenopus laevis (African clawed frog)
Molecular weightTheoretical: 129.009414 KDa
Recombinant expressionOrganism: Spodoptera frugiperda (fall armyworm)
SequenceString: SNAADGSQVF RFYWLDAYED QYSQPGVVYL FGKVWIESAD AYVSCCVSVK NIERTVYLLP RENRVQLSTG KDTGAPVSMM HVYQEFNEA VAEKYKIMKF KSKKVDKDYA FEIPDVPASS EYLEVRYSAD SPQLPQDLKG ETFSHVFGTN TSSLELFLLS R KIKGPSWL ...String:
SNAADGSQVF RFYWLDAYED QYSQPGVVYL FGKVWIESAD AYVSCCVSVK NIERTVYLLP RENRVQLSTG KDTGAPVSMM HVYQEFNEA VAEKYKIMKF KSKKVDKDYA FEIPDVPASS EYLEVRYSAD SPQLPQDLKG ETFSHVFGTN TSSLELFLLS R KIKGPSWL EIKSPQLSSQ PMSWCKVEAV VTRPDQVSVV KDLAPPPVVV LSLSMKTVQN AKTHQNEIVA IAALVHHTFP LD KAPPQPP FQTHFCVLSK LNDCIFPYDY NEAVKQKNAN IEIALTERTL LGFFLAKIHK IDPDVIVGHD IYGFDLEVLL QRI NSCKVP FWSKIGRLRR SVMPKLGGRS GFAERNAACG RIICDIEISA KELIRCKSYH LSELVHQILK AERVVIPPEN IRNA YNDSV HLLYMLENTW IDAKFILQIM CELNVLPLAL QITNIAGNVM SRTLMGGRSE RNEYLLLHAF TENNFIVPDK PVFKK MQQT TVEDNDDMGT DQNKNKSRKK AAYAGGLVLE PKVGFYDKFI LLLDFNSLYP SIIQEYNICF TTVHREAPST QKGEDQ DEI PELPHSDLEM GILPREIRKL VERRRHVKQL MKQPDLNPDL YLQYDIRQKA LKLTANSMYG CLGFSYSRFY AKPLAAL VT HQGREILLHT KEMVQKMNLE VIYGDTDSIM INTNCNNLEE VFKLGNRVKS EINKSYKLLE IDIDGIFKSL LLLKKKKY A ALTVEPTGDG KYVTKQELKG LDIVRRDWCE LAKQAGNYVI SQILSDQPRD SIVENIQKKL TEIGENVTNG TVPITQYEI NKALTKDPQD YPDKKSLPHV HVALWINSQG GRKVKAGDTI SYVICQDGSN LSASQRAYAQ EQLQKQENLS IDTQYYLSQQ VHPVVARIC EPIDGIDSAL IAMWLGLDPS QFRAHRHYQQ DEENDALLGG PSQLTDEEKY RDCERFKFFC PKCGTENIYD N VFDGSGLQ IEPGLKRCSK PECDASPLDY VIQVHNKLLL DIRRYIKKYY SGWLVCEEKT CQNRTRRLPL SFSRNGPICQ AC SKATLRS EYPEKALYTQ LCFYRFIFDW DYALEKVVSE QERGHLKKKL FQESENQYKK LKSTVDQVLS RSGYSEVNLS KLF QTLNTI K

UniProtKB: DNA polymerase alpha catalytic subunit

-
Macromolecule #2: DNA polymerase alpha subunit B

MacromoleculeName: DNA polymerase alpha subunit B / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
Molecular weightTheoretical: 67.194219 KDa
Recombinant expressionOrganism: Spodoptera frugiperda (fall armyworm)
SequenceString: SNAMSVSAKS IAEELKVFDV NFEDEEVPEK MVELCTVHRL KEEDMVNEWM AFSTTRNLPL TVGNLNLLEH EVLNKKSARP RPSLKKEKH CGNRDFNTIQ ELIEVETAEE NLLDSYATPA KGSQKRNLST PEHPQSKRIL SINRSPHVLF SPTSFSPSAT P SQKYGSRT ...String:
SNAMSVSAKS IAEELKVFDV NFEDEEVPEK MVELCTVHRL KEEDMVNEWM AFSTTRNLPL TVGNLNLLEH EVLNKKSARP RPSLKKEKH CGNRDFNTIQ ELIEVETAEE NLLDSYATPA KGSQKRNLST PEHPQSKRIL SINRSPHVLF SPTSFSPSAT P SQKYGSRT NRGEVVTTYG ELQGTTWNGG SGSNTNVELF TSLDEPLTKM YKFMFQKLMD IREVVSIKIE ELGASLKDHF QI DEFTSVS LPAQETVTVL GQIGCDSNGK LNSKSVILEG DREHSAGMQV PVDLSELKDY SLFPGQVVIM EGTNSTGRRF VPT KLYEGV PLPFHQPSKE FEECPQQMVI TACGPFTTSD TITYDALKDL IDIVNRDRPD ICILLGPFLD AKHEQIENLQ LTVT FEDVF KRCLKMIIEG TRPSGCHLVI VPSLRDVHHD PVYPQPPFSC FEPAKEDKER VHFVADPCTL SVNGVVIGMT STDLL FHMG AEEISSSAGA PDRFSRILRH ILTQRSYYPL YPPNEEINID YEALYSYTPM PVTPDVFIVP SELRYFIKDV TGCICI NPG RLTKGLVGGT YARFLVKSGA MGSEGKRSTC ISAQVVRV

UniProtKB: DNA polymerase alpha subunit B

-
Macromolecule #3: DNA primase large subunit

MacromoleculeName: DNA primase large subunit / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Xenopus laevis (African clawed frog)
Molecular weightTheoretical: 59.673844 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MLFSRDRKYR HNTRLTGDRK GDLYPSSLQF YQHPPTENIS LIEFETFAIE RLKLLKAVEN LGVSYVKNSE EYSKKLELEL RKLKFPYRP LHEEISDDVY DLRRKDHISH FILRLAYCQS EDLRRWFIQQ EMDLFKFRFG LLTKESVQEF LKLNDLQYVA I SEDEKNMH ...String:
MLFSRDRKYR HNTRLTGDRK GDLYPSSLQF YQHPPTENIS LIEFETFAIE RLKLLKAVEN LGVSYVKNSE EYSKKLELEL RKLKFPYRP LHEEISDDVY DLRRKDHISH FILRLAYCQS EDLRRWFIQQ EMDLFKFRFG LLTKESVQEF LKLNDLQYVA I SEDEKNMH KEDLMNSSFG LSLTKMEDTE FYKVPFQAAL DLVRPRKVFL WRGFAFIPHK DIVSIVLNDF RAKLSKALAL SA RSLPVVQ SDERLQPLLN HLSHSYIGQD FSSQSNTGKI SLEQIDGFAA KSFPLCMRQL HKSLRENHHL RHGGRMQYGL FLK GIGLTL EQALQFWRLE FTKGKVDSEK FDKVYAYSIR HNYGKEGKRT DYTPYSCMKV ILSNPPSQGD YHGCPFRHSD PELL KQKLQ SFKVPSSGIN QILELVKGMH YQLACQKYFE LTHSVDDCGF SLNHPNQYFA ESQKLLTGSR EIKKEQTARD SPAVT ASQL SSSSSSASIP KSQSSAPEME DLEQIFSEY

UniProtKB: DNA primase large subunit

-
Macromolecule #4: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 4 / Number of copies: 2 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

-
Macromolecule #5: IRON/SULFUR CLUSTER

MacromoleculeName: IRON/SULFUR CLUSTER / type: ligand / ID: 5 / Number of copies: 1 / Formula: SF4
Molecular weightTheoretical: 351.64 Da
Chemical component information

ChemComp-FS1:
IRON/SULFUR CLUSTER

-
Experimental details

-
Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

Concentration0.9 mg/mL
BufferpH: 7.5
VitrificationCryogen name: ETHANE

-
Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 1 / Number real images: 17820 / Average electron dose: 54.8 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 105000
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

+
Image processing

Particle selectionNumber selected: 23007566
Startup modelType of model: INSILICO MODEL / In silico model: Ab initio reconstruction
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 2.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Number images used: 334066
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
FSC plot (resolution estimation)

-
Atomic model buiding 1

Initial modelPDB ID:

5exr


Chain - Source name: SwissModel / Chain - Initial model type: in silico model
RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-8g99:
Partial auto-inhibitory complex of Xenopus laevis DNA polymerase alpha-primase

+
About Yorodumi

-
News

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

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242)：Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more