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- EMDB-19332: DNA helicase RECQL5 in complex with homo Di-Gluebody G5-006 - REC... -

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Basic information

Entry
Database: EMDB / ID: EMD-19332
TitleDNA helicase RECQL5 in complex with homo Di-Gluebody G5-006 - RECQL5 local refinement
Map dataEMhancer: RECQL5 in complex with G5-006 Di-Gluebody: local refinement of RECQL5
Sample
  • Complex: Local refinement of the RECQL5 part of RECQL5 homodimer assembled by G5-006 homo Di-Gluebody
    • Protein or peptide: ATP-dependent DNA helicase Q5
  • Ligand: ZINC ION
KeywordsDNA helicase / Di-Gluebody / HYDROLASE
Function / homology
Function and homology information


mitotic DNA-templated DNA replication / chromosome separation / cellular response to camptothecin / replication-born double-strand break repair via sister chromatid exchange / transcription preinitiation complex / 3'-5' DNA helicase activity / DNA 3'-5' helicase / DNA metabolic process / RNA polymerase II complex binding / negative regulation of transcription elongation by RNA polymerase II ...mitotic DNA-templated DNA replication / chromosome separation / cellular response to camptothecin / replication-born double-strand break repair via sister chromatid exchange / transcription preinitiation complex / 3'-5' DNA helicase activity / DNA 3'-5' helicase / DNA metabolic process / RNA polymerase II complex binding / negative regulation of transcription elongation by RNA polymerase II / negative regulation of double-strand break repair via homologous recombination / DNA helicase activity / replication fork / isomerase activity / helicase activity / double-strand break repair via homologous recombination / cellular response to xenobiotic stimulus / mitotic cell cycle / chromosome / forked DNA-dependent helicase activity / single-stranded 3'-5' DNA helicase activity / four-way junction helicase activity / double-stranded DNA helicase activity / DNA replication / cell division / DNA repair / ATP hydrolysis activity / DNA binding / nucleoplasm / ATP binding / metal ion binding / identical protein binding / nucleus / cytosol / cytoplasm
Similarity search - Function
RecQ helicase-like 5 / RecQ helicase protein-like 5 (RecQ5) / Set2 Rpb1 interacting domain / SRI (Set2 Rpb1 interacting) domain / ATP-dependent DNA helicase RecQ, zinc-binding domain / RecQ zinc-binding / DNA helicase, ATP-dependent, RecQ type / DNA/RNA helicase, ATP-dependent, DEAH-box type, conserved site / DEAH-box subfamily ATP-dependent helicases signature. / DEAD/DEAH box helicase domain ...RecQ helicase-like 5 / RecQ helicase protein-like 5 (RecQ5) / Set2 Rpb1 interacting domain / SRI (Set2 Rpb1 interacting) domain / ATP-dependent DNA helicase RecQ, zinc-binding domain / RecQ zinc-binding / DNA helicase, ATP-dependent, RecQ type / DNA/RNA helicase, ATP-dependent, DEAH-box type, conserved site / DEAH-box subfamily ATP-dependent helicases signature. / DEAD/DEAH box helicase domain / DEAD/DEAH box helicase / Helicase conserved C-terminal domain / helicase superfamily c-terminal domain / Superfamilies 1 and 2 helicase C-terminal domain profile. / Superfamilies 1 and 2 helicase ATP-binding type-1 domain profile. / DEAD-like helicases superfamily / Helicase, C-terminal / Helicase superfamily 1/2, ATP-binding domain / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
ATP-dependent DNA helicase Q5
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.18 Å
AuthorsYi G / Ye M / Mamalis D / Sauer DB / von Delft F / Davis BG / Gilbert RJC
Funding support United Kingdom, 1 items
OrganizationGrant numberCountry
Other private United Kingdom
CitationJournal: To Be Published
Title: Di-Gluebodies: Rigid modular nanobody protein assemblies enabling simultaneous determination of high-resolution cryo-EM structures
Authors: Yi G / Mamalis D / Ye M / Carrique L / Fairhead M / Li H / Duerr K / Zhang P / Sauer DB / von Delft F / Davis BG / Gilbert RJC
History
DepositionJan 2, 2024-
Header (metadata) releaseJan 15, 2025-
Map releaseJan 15, 2025-
UpdateJan 15, 2025-
Current statusJan 15, 2025Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_19332.png

Downloads & links

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Map

FileDownload / File: emd_19332.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationEMhancer: RECQL5 in complex with G5-006 Di-Gluebody: local refinement of RECQL5
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.7 Å/pix.
x 420 pix.
= 294. Å		0.7 Å/pix.
x 420 pix.
= 294. Å		0.7 Å/pix.
x 420 pix.
= 294. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.7 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.26
Minimum - Maximum-0.029755948 - 1.9453092
Average (Standard dev.)0.00042445332 (±0.01176735)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions420420420
Spacing420420420
CellA=B=C: 294.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_19332_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: RECQL5 in complex with G5-006 Di-Gluebody: local refinement of RECQL5

Fileemd_19332_additional_1.map
AnnotationRECQL5 in complex with G5-006 Di-Gluebody: local refinement of RECQL5
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map A

Fileemd_19332_half_map_1.map
Annotationhalf map A
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map B

Fileemd_19332_half_map_2.map
Annotationhalf map B
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Local refinement of the RECQL5 part of RECQL5 homodimer assembled...

EntireName: Local refinement of the RECQL5 part of RECQL5 homodimer assembled by G5-006 homo Di-Gluebody
Components
  • Complex: Local refinement of the RECQL5 part of RECQL5 homodimer assembled by G5-006 homo Di-Gluebody
    • Protein or peptide: ATP-dependent DNA helicase Q5
  • Ligand: ZINC ION

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Supramolecule #1: Local refinement of the RECQL5 part of RECQL5 homodimer assembled...

SupramoleculeName: Local refinement of the RECQL5 part of RECQL5 homodimer assembled by G5-006 homo Di-Gluebody
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 126 KDa

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Macromolecule #1: ATP-dependent DNA helicase Q5

MacromoleculeName: ATP-dependent DNA helicase Q5 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 49.20382 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: PERRVRSTLK KVFGFDSFKT PLQESATMAV VKGNKDVFVC MPTGAGKSLC YQLPALLAKG ITIVVSPLIA LIQDQVDHLL TLKVRVSSL NSKLSAQERK ELLADLEREK PQTKILYITP EMAASSSFQP TLNSLVSRHL LSYLVVDEAH CVSQWGHDFR P DYLRLGAL ...String:
PERRVRSTLK KVFGFDSFKT PLQESATMAV VKGNKDVFVC MPTGAGKSLC YQLPALLAKG ITIVVSPLIA LIQDQVDHLL TLKVRVSSL NSKLSAQERK ELLADLEREK PQTKILYITP EMAASSSFQP TLNSLVSRHL LSYLVVDEAH CVSQWGHDFR P DYLRLGAL RSRLGHAPCV ALTATATPQV QEDVFAALHL KKPVAIFKTP CFRANLFYDV QFKELISDPY GNLKDFCLKA LG QEADKGL SGCGIVYCRT REACEQLAIE LSCRGVNAKA YHAGLKASER TLVQNDWMEE KVPVIVATIS FGMGVDKANV RFV AHWNIA KSMAGYYQES GRAGRDGKPS WCRLYYSRND RDQVSFLIRK EVAKLQEKRG NKASDKATIM AFDALVTFCE ELGC RHAAI AKYFGDALPA CAKGCDHCQN PTAVRRRLEA LERSSSW

UniProtKB: ATP-dependent DNA helicase Q5

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Macromolecule #2: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 2 / Number of copies: 1 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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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

Concentration0.8 mg/mL
BufferpH: 7.5
Component:
ConcentrationFormulaName
20.0 mMC8H18N2O4SHEPES
150.0 mMNaClsodium chloride
GridModel: C-flat-2/1 / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Number real images: 9247 / Average electron dose: 60.0 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: 3.0 µm / Nominal defocus min: 1.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 610675
Startup modelType of model: INSILICO MODEL / In silico model: Ab-initio reconstruction in cryosparc
Final reconstructionNumber classes used: 4 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.18 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.0) / Number images used: 194417
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.0) / Details: Non-uniform refinement in Cryosparc
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.0) / Details: Local refinement in Cryosparc
Final 3D classificationNumber classes: 6 / Software - Name: cryoSPARC (ver. 4.0)
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

7zmv


Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Overall B value: 157.2
Output model

PDB-8rl6:
DNA helicase RECQL5 in complex with homo Di-Gluebody G5-006 - RECQL5 local refinement

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