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Yorodumi- EMDB-44310: Swc3-Swc2 subcomplex from SWR1-nucleosome complex (focused refinement) -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-44310 | ||||||||||||
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Title | Swc3-Swc2 subcomplex from SWR1-nucleosome complex (focused refinement) | ||||||||||||
Map data | |||||||||||||
Sample |
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Keywords | Chromatin Remodeler / Snf2 family ATPase / histone exchange / H2A.Z / GENE REGULATION | ||||||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 7.3 Å | ||||||||||||
Authors | Louder RK / Park G / Wu C | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: Cell / Year: 2024 Title: Molecular basis of global promoter sensing and nucleosome capture by the SWR1 chromatin remodeler. Authors: Robert K Louder / Giho Park / Ziyang Ye / Justin S Cha / Anne M Gardner / Qin Lei / Anand Ranjan / Eva Höllmüller / Florian Stengel / B Franklin Pugh / Carl Wu / Abstract: The SWR1 chromatin remodeling complex is recruited to +1 nucleosomes downstream of transcription start sites of eukaryotic promoters, where it exchanges histone H2A for the specialized variant H2A.Z. ...The SWR1 chromatin remodeling complex is recruited to +1 nucleosomes downstream of transcription start sites of eukaryotic promoters, where it exchanges histone H2A for the specialized variant H2A.Z. Here, we use cryoelectron microscopy (cryo-EM) to resolve the structural basis of the SWR1 interaction with free DNA, revealing a distinct open conformation of the Swr1 ATPase that enables sliding from accessible DNA to nucleosomes. A complete structural model of the SWR1-nucleosome complex illustrates critical roles for Swc2 and Swc3 subunits in oriented nucleosome engagement by SWR1. Moreover, an extended DNA-binding α helix within the Swc3 subunit enables sensing of nucleosome linker length and is essential for SWR1-promoter-specific recruitment and activity. The previously unresolved N-SWR1 subcomplex forms a flexible extended structure, enabling multivalent recognition of acetylated histone tails by reader domains to further direct SWR1 toward the +1 nucleosome. Altogether, our findings provide a generalizable mechanism for promoter-specific targeting of chromatin and transcription complexes. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_44310.map.gz | 2.3 MB | EMDB map data format | |
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Header (meta data) | emd-44310-v30.xml emd-44310.xml | 16.7 KB 16.7 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_44310_fsc.xml | 13.7 KB | Display | FSC data file |
Images | emd_44310.png | 70.8 KB | ||
Masks | emd_44310_msk_1.map | 216 MB | Mask map | |
Filedesc metadata | emd-44310.cif.gz | 4.6 KB | ||
Others | emd_44310_half_map_1.map.gz emd_44310_half_map_2.map.gz | 171.3 MB 171.2 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-44310 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-44310 | HTTPS FTP |
-Validation report
Summary document | emd_44310_validation.pdf.gz | 628.2 KB | Display | EMDB validaton report |
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Full document | emd_44310_full_validation.pdf.gz | 627.8 KB | Display | |
Data in XML | emd_44310_validation.xml.gz | 21 KB | Display | |
Data in CIF | emd_44310_validation.cif.gz | 27.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-44310 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-44310 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_44310.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.03 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_44310_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_44310_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_44310_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Native SWR1 bound to nucleosome
Entire | Name: Native SWR1 bound to nucleosome |
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Components |
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-Supramolecule #1: Native SWR1 bound to nucleosome
Supramolecule | Name: Native SWR1 bound to nucleosome / type: complex / ID: 1 / Parent: 0 Details: Endogenously purified yeast SWR1 complex bound to 60-N-7 nucleosome fragment in the presence of ADP-BeF3. |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: W303 |
Molecular weight | Theoretical: 1.19 MDa |
-Supramolecule #2: Native SWR1 complex
Supramolecule | Name: Native SWR1 complex / type: complex / ID: 2 / Parent: 1 / Details: Endogenously purified yeast SWR1 complex |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: W303 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.08 mg/mL |
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Buffer | pH: 7.6 Details: 80 nM SWR1, 160 nM nucleosomes, 1 mM ADP, 10 mM NaF, 8 mM BeCl2, 0.05% glutaraldehyde, 20 mM HEPES-KOH pH 7.6, 1.5 mM MgCl2, 0.25 mM TCEP, 0.01% IGEPAL CA-630, 1% glycerol |
Grid | Model: Quantifoil / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 90 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: 3 second blot time and blot force of 10.. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 7260 / Average exposure time: 4.0 sec. / Average electron dose: 54.0 e/Å2 Details: Each micrograph was fractionated into 64 frames within a 4 second exposure. |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Calibrated magnification: 48543 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.6 µm / Nominal defocus min: 2.0 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |