[English] 日本語

- EMDB-3243: Dodecameric Chaetomium thermophilum Rvb1/Rvb2 complex in stretche... -
+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-3243 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Dodecameric Chaetomium thermophilum Rvb1/Rvb2 complex in stretched conformation | |||||||||
![]() | Cryo-EM reconstruction of Chaetomium thermophilum dodecameric Rvb1/Rvb2 complex in stretched conformation | |||||||||
![]() |
| |||||||||
![]() | AAA+ proteins / cryo-electron microscopy / Rvb1 / Rvb2 / Chaetomium thermophilum | |||||||||
Function / homology | ![]() R2TP complex / Swr1 complex / Ino80 complex / box C/D snoRNP assembly / NuA4 histone acetyltransferase complex / DNA helicase activity / DNA helicase / chromatin remodeling / DNA repair / regulation of transcription by RNA polymerase II ...R2TP complex / Swr1 complex / Ino80 complex / box C/D snoRNP assembly / NuA4 histone acetyltransferase complex / DNA helicase activity / DNA helicase / chromatin remodeling / DNA repair / regulation of transcription by RNA polymerase II / ATP hydrolysis activity / ATP binding Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 21.4 Å | |||||||||
![]() | Silva-Martin N / Dauden MI / Glatt S / Hoffmann NA / Kastritis P / Bork P / Beck M / Mueller CW | |||||||||
![]() | ![]() Title: The Combination of X-Ray Crystallography and Cryo-Electron Microscopy Provides Insight into the Overall Architecture of the Dodecameric Rvb1/Rvb2 Complex. Authors: Noella Silva-Martin / María I Daudén / Sebastian Glatt / Niklas A Hoffmann / Panagiotis Kastritis / Peer Bork / Martin Beck / Christoph W Müller / ![]() Abstract: The Rvb1/Rvb2 complex is an essential component of many cellular pathways. The Rvb1/Rvb2 complex forms a dodecameric assembly where six copies of each subunit form two heterohexameric rings. However, ...The Rvb1/Rvb2 complex is an essential component of many cellular pathways. The Rvb1/Rvb2 complex forms a dodecameric assembly where six copies of each subunit form two heterohexameric rings. However, due to conformational variability, the way the two rings pack together is still not fully understood. Here, we present the crystal structure and two cryo-electron microscopy reconstructions of the dodecameric, full-length Rvb1/Rvb2 complex, all showing that the interaction between the two heterohexameric rings is mediated through the Rvb1/Rvb2-specific domain II. Two conformations of the Rvb1/Rvb2 dodecamer are present in solution: a stretched conformation also present in the crystal, and a compact conformation. Novel asymmetric features observed in the reconstruction of the compact conformation provide additional insight into the plasticity of the Rvb1/Rvb2 complex. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 1.1 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 11.2 KB 11.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 2.6 KB | Display | ![]() |
Images | ![]() | 45.7 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 248.3 KB | Display | ![]() |
---|---|---|---|---|
Full document | ![]() | 247.4 KB | Display | |
Data in XML | ![]() | 6.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
-
Links
EMDB pages | ![]() ![]() |
---|
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Cryo-EM reconstruction of Chaetomium thermophilum dodecameric Rvb1/Rvb2 complex in stretched conformation | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 4.34 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
-Entire : Full-length Chaetomium thermophilum Rvb1/Rvb2 dodecameric complex...
Entire | Name: Full-length Chaetomium thermophilum Rvb1/Rvb2 dodecameric complex in stretched conformation |
---|---|
Components |
|
-Supramolecule #1000: Full-length Chaetomium thermophilum Rvb1/Rvb2 dodecameric complex...
Supramolecule | Name: Full-length Chaetomium thermophilum Rvb1/Rvb2 dodecameric complex in stretched conformation type: sample / ID: 1000 Oligomeric state: Dodecamer formed by two hetero-hexamers of Rvb1/Rvb2 Number unique components: 2 |
---|---|
Molecular weight | Theoretical: 621 KDa |
-Macromolecule #1: Rvb1
Macromolecule | Name: Rvb1 / type: protein_or_peptide / ID: 1 / Name.synonym: RuvBL1, Pontin, TIP49a / Number of copies: 6 / Oligomeric state: Double hetero-hexamer with Rvb2 / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 50.384 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: RuvB-like helicase InterPro: AAA+ ATPase domain, P-loop containing nucleoside triphosphate hydrolase, RuvB-like, TIP49, P-loop domain |
-Macromolecule #2: Rvb2
Macromolecule | Name: Rvb2 / type: protein_or_peptide / ID: 2 / Name.synonym: RuvBL2, Reptin, TIP49b / Number of copies: 6 / Oligomeric state: Double hetero-hexamer with Rvb2 / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 53.1458 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: RuvB-like helicase InterPro: AAA+ ATPase domain, P-loop containing nucleoside triphosphate hydrolase, RuvB-like, TIP49, P-loop domain |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | single particle reconstruction |
Aggregation state | particle |
-
Sample preparation
Concentration | 0.35 mg/mL |
---|---|
Buffer | pH: 7.5 Details: 20 mM Tris-HCl pH 7.5, 125 mM NaCl, 2 mM 2-mercaptoethanol and 5 mM MgCl2 |
Grid | Details: glow-discharged molybdenum grids 400 mesh, 1.2/1.5, Quantifoil |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 120 K / Instrument: FEI VITROBOT MARK III Method: 9 seconds blotting time, 15 seconds waiting time, blot offset -3 |
-
Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Date | Nov 3, 2014 |
Image recording | Category: CCD / Film or detector model: FEI FALCON II (4k x 4k) / Number real images: 1629 / Average electron dose: 40 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 4.2 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 75000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |