+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-15187 | ||||||||||||||||||
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Title | A-module of Chaetomium thermophilum INO80 bound to curved DNA | ||||||||||||||||||
Map data | main map | ||||||||||||||||||
Sample |
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Biological species | Thermochaetoides thermophila (fungus) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | ||||||||||||||||||
Authors | Kunert F / Metzner FJ / Eustermann S / Jung J / Woike S / Schall K / Kostrewa D / Hopfner KP | ||||||||||||||||||
Funding support | European Union, Germany, 5 items
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Citation | Journal: Sci Adv / Year: 2022 Title: Structural mechanism of extranucleosomal DNA readout by the INO80 complex. Authors: Franziska Kunert / Felix J Metzner / James Jung / Markus Höpfler / Stephan Woike / Kevin Schall / Dirk Kostrewa / Manuela Moldt / Jia-Xuan Chen / Susanne Bantele / Boris Pfander / Sebastian ...Authors: Franziska Kunert / Felix J Metzner / James Jung / Markus Höpfler / Stephan Woike / Kevin Schall / Dirk Kostrewa / Manuela Moldt / Jia-Xuan Chen / Susanne Bantele / Boris Pfander / Sebastian Eustermann / Karl-Peter Hopfner / Abstract: The nucleosomal landscape of chromatin depends on the concerted action of chromatin remodelers. The INO80 remodeler specifically places nucleosomes at the boundary of gene regulatory elements, which ...The nucleosomal landscape of chromatin depends on the concerted action of chromatin remodelers. The INO80 remodeler specifically places nucleosomes at the boundary of gene regulatory elements, which is proposed to be the result of an ATP-dependent nucleosome sliding activity that is regulated by extranucleosomal DNA features. Here, we use cryo-electron microscopy and functional assays to reveal how INO80 binds and is regulated by extranucleosomal DNA. Structures of the regulatory A-module bound to DNA clarify the mechanism of linker DNA binding. The A-module is connected to the motor unit via an HSA/post-HSA lever element to chemomechanically couple the motor and linker DNA sensing. Two notable sites of curved DNA recognition by coordinated action of the four actin/actin-related proteins and the motor suggest how sliding by INO80 can be regulated by extranucleosomal DNA features. Last, the structures clarify the recruitment of YY1/Ies4 subunits and reveal deep architectural similarities between the regulatory modules of INO80 and SWI/SNF complexes. | ||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_15187.map.gz | 49.7 MB | EMDB map data format | |
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Header (meta data) | emd-15187-v30.xml emd-15187.xml | 14.5 KB 14.5 KB | Display Display | EMDB header |
Images | emd_15187.png | 73 KB | ||
Others | emd_15187_half_map_1.map.gz emd_15187_half_map_2.map.gz | 49.7 MB 49.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-15187 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-15187 | HTTPS FTP |
-Validation report
Summary document | emd_15187_validation.pdf.gz | 836.2 KB | Display | EMDB validaton report |
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Full document | emd_15187_full_validation.pdf.gz | 835.8 KB | Display | |
Data in XML | emd_15187_validation.xml.gz | 12.1 KB | Display | |
Data in CIF | emd_15187_validation.cif.gz | 14.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15187 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15187 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_15187.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | main map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.059 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: half map A
File | emd_15187_half_map_1.map | ||||||||||||
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Annotation | half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map B
File | emd_15187_half_map_2.map | ||||||||||||
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Annotation | half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : INO80 A-Module (Arp4-Ies4-N-actin-Arp8-Ino80HSA) on curved DNA
Entire | Name: INO80 A-Module (Arp4-Ies4-N-actin-Arp8-Ino80HSA) on curved DNA |
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Components |
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-Supramolecule #1: INO80 A-Module (Arp4-Ies4-N-actin-Arp8-Ino80HSA) on curved DNA
Supramolecule | Name: INO80 A-Module (Arp4-Ies4-N-actin-Arp8-Ino80HSA) on curved DNA type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: #1-#7 |
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Source (natural) | Organism: Thermochaetoides thermophila (fungus) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 42.4 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.9 µm / Nominal defocus min: 1.1 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Details | Ino80 A-module (Arp8, N-Actin, Arp4, Ies4) assembled on the HSA domain and bound to curved DNA |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.7 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 29000 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |