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Yorodumi- EMDB-21484: Cryo-EM structure of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosom... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21484 | |||||||||
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Title | Cryo-EM structure of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome in ADP Beryllium Fluoride state | |||||||||
Map data | cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome; unsharpened map | |||||||||
Sample |
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Keywords | Chromatin remodeling / Nucleosome / Gene Regulation / MOTOR PROTEIN | |||||||||
Function / homology | Function and homology information RHO GTPases activate IQGAPs / RHO GTPases Activate WASPs and WAVEs / Regulation of actin dynamics for phagocytic cup formation / chromatin remodeling at centromere / Platelet degranulation / DNA translocase activity / RSC-type complex / SWI/SNF complex / nucleosome disassembly / ATP-dependent chromatin remodeler activity ...RHO GTPases activate IQGAPs / RHO GTPases Activate WASPs and WAVEs / Regulation of actin dynamics for phagocytic cup formation / chromatin remodeling at centromere / Platelet degranulation / DNA translocase activity / RSC-type complex / SWI/SNF complex / nucleosome disassembly / ATP-dependent chromatin remodeler activity / NuA4 histone acetyltransferase complex / chromosome, centromeric region / ATP-dependent activity, acting on DNA / cytoskeleton organization / histone reader activity / meiotic cell cycle / chromosome segregation / helicase activity / transcription elongation by RNA polymerase II / lysine-acetylated histone binding / base-excision repair / structural constituent of chromatin / nucleosome / double-strand break repair / nucleosome assembly / chromatin organization / DNA helicase / chromatin remodeling / protein heterodimerization activity / chromatin binding / regulation of DNA-templated transcription / chromatin / regulation of transcription by RNA polymerase II / structural molecule activity / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / DNA binding / ATP binding / nucleus Similarity search - Function | |||||||||
Biological species | Xenopus laevis (African clawed frog) / synthetic construct (others) / Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) / Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||
Authors | Leschziner AE / Baker RW | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2021 Title: Structural insights into assembly and function of the RSC chromatin remodeling complex. Authors: Richard W Baker / Janice M Reimer / Peter J Carman / Bengi Turegun / Tsutomu Arakawa / Roberto Dominguez / Andres E Leschziner / Abstract: SWI/SNF chromatin remodelers modify the position and spacing of nucleosomes and, in humans, are linked to cancer. To provide insights into the assembly and regulation of this protein family, we ...SWI/SNF chromatin remodelers modify the position and spacing of nucleosomes and, in humans, are linked to cancer. To provide insights into the assembly and regulation of this protein family, we focused on a subcomplex of the Saccharomyces cerevisiae RSC comprising its ATPase (Sth1), the essential actin-related proteins (ARPs) Arp7 and Arp9 and the ARP-binding protein Rtt102. Cryo-EM and biochemical analyses of this subcomplex shows that ARP binding induces a helical conformation in the helicase-SANT-associated (HSA) domain of Sth1. Surprisingly, the ARP module is rotated 120° relative to the full RSC about a pivot point previously identified as a regulatory hub in Sth1, suggesting that large conformational changes are part of Sth1 regulation and RSC assembly. We also show that a conserved interaction between Sth1 and the nucleosome acidic patch enhances remodeling. As some cancer-associated mutations dysregulate rather than inactivate SWI/SNF remodelers, our insights into RSC complex regulation advance a mechanistic understanding of chromatin remodeling in disease states. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_21484.map.gz | 80.5 MB | EMDB map data format | |
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Header (meta data) | emd-21484-v30.xml emd-21484.xml | 61.2 KB 61.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_21484_fsc.xml | 10.7 KB | Display | FSC data file |
Images | emd_21484.png | 59.6 KB | ||
Filedesc metadata | emd-21484.cif.gz | 9.6 KB | ||
Others | emd_21484_additional_1.map.gz emd_21484_additional_10.map.gz emd_21484_additional_11.map.gz emd_21484_additional_12.map.gz emd_21484_additional_13.map.gz emd_21484_additional_2.map.gz emd_21484_additional_3.map.gz emd_21484_additional_4.map.gz emd_21484_additional_5.map.gz emd_21484_additional_6.map.gz emd_21484_additional_7.map.gz emd_21484_additional_8.map.gz emd_21484_additional_9.map.gz emd_21484_half_map_1.map.gz emd_21484_half_map_2.map.gz | 96 MB 60.4 MB 60.3 MB 95.1 MB 60.3 MB 73.8 MB 73.2 MB 73.8 MB 95.6 MB 71.9 MB 72.3 MB 72.3 MB 95.8 MB 80.8 MB 80.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21484 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21484 | HTTPS FTP |
-Validation report
Summary document | emd_21484_validation.pdf.gz | 732.9 KB | Display | EMDB validaton report |
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Full document | emd_21484_full_validation.pdf.gz | 732.5 KB | Display | |
Data in XML | emd_21484_validation.xml.gz | 17.9 KB | Display | |
Data in CIF | emd_21484_validation.cif.gz | 23.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21484 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21484 | HTTPS FTP |
-Related structure data
Related structure data | 6vz4MC 6vzgC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_21484.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome; unsharpened map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.16 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Additional map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Half map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
+Half map: cryo-EM map of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosome;...
-Sample components
+Entire : cryo-EM structure of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosom...
+Supramolecule #1: cryo-EM structure of Sth1-Arp7-Arp9-Rtt102 bound to the nucleosom...
+Supramolecule #2: Histone H3, Histone H4, Histone H2A, Histone H2B
+Supramolecule #3: DNA
+Supramolecule #4: Nuclear protein STH1/NPS1, Actin-related protein 7, Actin-like pr...
+Macromolecule #1: Histone H3
+Macromolecule #2: Histone H4
+Macromolecule #3: Histone H2A
+Macromolecule #4: Histone H2B
+Macromolecule #7: Nuclear protein STH1/NPS1
+Macromolecule #8: Actin-related protein 7
+Macromolecule #9: Actin-like protein ARP9
+Macromolecule #10: Regulator of Ty1 transposition protein 102
+Macromolecule #5: DNA (185-MER)
+Macromolecule #6: DNA (185-MER)
+Macromolecule #11: MAGNESIUM ION
+Macromolecule #12: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #13: BERYLLIUM TRIFLUORIDE ION
+Macromolecule #14: ADENOSINE-5'-TRIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 1 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: UltrAuFoil / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR / Details: Glow discharge for 30 seconds at 25 mAmp |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: 4 second blot time, blot force 20. |
Details | Sample was crosslinked using the GRAFIX protocol |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average exposure time: 7.0 sec. / Average electron dose: 53.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 36000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model |
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Details | Initial model docking was done in Chimera. Phenix.real_space_refine was used to refine the nucleosome model. Sth1-Arp7-Arp9-Rtt102 were refined in Rosetta and the top ten models were deposited. | ||||||||
Refinement | Space: REAL / Protocol: FLEXIBLE FIT | ||||||||
Output model | PDB-6vz4: |