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Yorodumi- EMDB-52442: Focused refined map of PAF complex in the EC-DSIF-PAF-SPT6-RECQL5... -
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Open data
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Basic information
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| Title | Focused refined map of PAF complex in the EC-DSIF-PAF-SPT6-RECQL5 complex | |||||||||
Map data | Sharpened focused refined map of PAF | |||||||||
Sample |
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Keywords | transcription elongation / DNA helicase / transcription-coupled repair / RNA polymerase II / TRANSCRIPTION | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.5 Å | |||||||||
Authors | Zhang L / Zhang S | |||||||||
| Funding support | United Kingdom, 1 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2025Title: Structural basis of RECQL5-induced RNA polymerase II transcription braking and subsequent reactivation. Authors: Luojia Zhang / Yuliya Gordiyenko / Tomos Morgan / Catarina Franco / Ana Tufegdžić Vidaković / Suyang Zhang / ![]() Abstract: Abnormally fast transcription elongation can lead to detrimental consequences such as transcription-replication collisions, altered alternative splicing patterns and genome instability. Therefore, ...Abnormally fast transcription elongation can lead to detrimental consequences such as transcription-replication collisions, altered alternative splicing patterns and genome instability. Therefore, elongating RNA polymerase II (Pol II) requires mechanisms to slow its progression, yet the molecular basis of transcription braking remains unclear. RECQL5 is a DNA helicase that functions as a general elongation factor by slowing down Pol II. Here we report cryo-electron microscopy structures of human RECQL5 bound to multiple transcription elongation complexes. Combined with biochemical analysis, we identify an α-helix of RECQL5 responsible for binding Pol II and slowdown of transcription elongation. We further reveal that the transcription-coupled DNA repair (TCR) complex allows Pol II to overcome RECQL5-induced transcription braking through concerted actions of its translocase activity and competition with RECQL5 for engaging Pol II. Additionally, RECQL5 inhibits TCR-mediated Pol II ubiquitination to prevent activation of the DNA repair pathway. Our results suggest a model in which RECQL5 and the TCR complex coordinately regulate transcription elongation rates to ensure transcription efficiency while maintaining genome stability. | |||||||||
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Structure visualization
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_52442.map.gz | 168.1 MB | EMDB map data format | |
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| Header (meta data) | emd-52442-v30.xml emd-52442.xml | 16.2 KB 16.2 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_52442_fsc.xml | 11.8 KB | Display | FSC data file |
| Images | emd_52442.png | 76.6 KB | ||
| Masks | emd_52442_msk_1.map | 178 MB | Mask map | |
| Filedesc metadata | emd-52442.cif.gz | 4.2 KB | ||
| Others | emd_52442_additional_1.map.gz emd_52442_half_map_1.map.gz emd_52442_half_map_2.map.gz | 89.7 MB 164.9 MB 164.9 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-52442 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-52442 | HTTPS FTP |
-Validation report
| Summary document | emd_52442_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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| Full document | emd_52442_full_validation.pdf.gz | 1.1 MB | Display | |
| Data in XML | emd_52442_validation.xml.gz | 20.8 KB | Display | |
| Data in CIF | emd_52442_validation.cif.gz | 26.9 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-52442 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-52442 | HTTPS FTP |
-Related structure data
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_52442.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Sharpened focused refined map of PAF | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.8156 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Mask #1
| File | emd_52442_msk_1.map | ||||||||||||
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| Density Histograms |
-Additional map: Non-sharpened focused refined map of PAF
| File | emd_52442_additional_1.map | ||||||||||||
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| Annotation | Non-sharpened focused refined map of PAF | ||||||||||||
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| Density Histograms |
-Half map: Half map for the focused refined map of PAF
| File | emd_52442_half_map_1.map | ||||||||||||
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| Annotation | Half map for the focused refined map of PAF | ||||||||||||
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| Density Histograms |
-Half map: Half map for the focused refined map of PAF
| File | emd_52442_half_map_2.map | ||||||||||||
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| Annotation | Half map for the focused refined map of PAF | ||||||||||||
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| Density Histograms |
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Sample components
-Entire : Pol II-DSIF-PAF-SPT6-RECQL5 complex
| Entire | Name: Pol II-DSIF-PAF-SPT6-RECQL5 complex |
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| Components |
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-Supramolecule #1: Pol II-DSIF-PAF-SPT6-RECQL5 complex
| Supramolecule | Name: Pol II-DSIF-PAF-SPT6-RECQL5 complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 |
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| Source (natural) | Organism: Homo sapiens (human) |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.2 mg/mL |
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| Buffer | pH: 7.5 Details: 20 mM HEPES pH 7.5, 50 mM KCl, 4 mM MgCl2, 1 mM DTT |
| Grid | Model: Quantifoil R3.5/1 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 0.25 |
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Number grids imaged: 2 / Number real images: 71964 / Average electron dose: 40.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 96000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi



Keywords
Homo sapiens (human)
Authors
United Kingdom, 1 items
Citation






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Processing
FIELD EMISSION GUN


