+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-4151 | |||||||||
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Title | S.cerevisiae Elp123 sub-complex | |||||||||
Map data | Negative staining reconstruction of the S.cerevisiae Elp123 sub-complex | |||||||||
Sample |
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Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 27.0 Å | |||||||||
Authors | Dauden MI / Kosinski J / Kolaj-Robin O / Desfosses A / Ori A / Faux C / Hoffmann NA / Onuma OF / Breuring KD / Beck M ...Dauden MI / Kosinski J / Kolaj-Robin O / Desfosses A / Ori A / Faux C / Hoffmann NA / Onuma OF / Breuring KD / Beck M / Sachse C / Seraphin B / Glatt S / Mueller CW | |||||||||
Citation | Journal: EMBO Rep / Year: 2017 Title: Architecture of the yeast Elongator complex. Authors: Maria I Dauden / Jan Kosinski / Olga Kolaj-Robin / Ambroise Desfosses / Alessandro Ori / Celine Faux / Niklas A Hoffmann / Osita F Onuma / Karin D Breunig / Martin Beck / Carsten Sachse / ...Authors: Maria I Dauden / Jan Kosinski / Olga Kolaj-Robin / Ambroise Desfosses / Alessandro Ori / Celine Faux / Niklas A Hoffmann / Osita F Onuma / Karin D Breunig / Martin Beck / Carsten Sachse / Bertrand Séraphin / Sebastian Glatt / Christoph W Müller / Abstract: The highly conserved eukaryotic Elongator complex performs specific chemical modifications on wobble base uridines of tRNAs, which are essential for proteome stability and homeostasis. The complex is ...The highly conserved eukaryotic Elongator complex performs specific chemical modifications on wobble base uridines of tRNAs, which are essential for proteome stability and homeostasis. The complex is formed by six individual subunits (Elp1-6) that are all equally important for its tRNA modification activity. However, its overall architecture and the detailed reaction mechanism remain elusive. Here, we report the structures of the fully assembled yeast Elongator and the Elp123 sub-complex solved by an integrative structure determination approach showing that two copies of the Elp1, Elp2, and Elp3 subunits form a two-lobed scaffold, which binds Elp456 asymmetrically. Our topological models are consistent with previous studies on individual subunits and further validated by complementary biochemical analyses. Our study provides a structural framework on how the tRNA modification activity is carried out by Elongator. | |||||||||
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_4151.map.gz | 32 MB | EMDB map data format | |
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Header (meta data) | emd-4151-v30.xml emd-4151.xml | 12.2 KB 12.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_4151_fsc.xml | 9.4 KB | Display | FSC data file |
Images | emd_4151.png | 65.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-4151 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4151 | HTTPS FTP |
-Validation report
Summary document | emd_4151_validation.pdf.gz | 237.5 KB | Display | EMDB validaton report |
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Full document | emd_4151_full_validation.pdf.gz | 236.6 KB | Display | |
Data in XML | emd_4151_validation.xml.gz | 10.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4151 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4151 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_4151.map.gz / Format: CCP4 / Size: 42.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Negative staining reconstruction of the S.cerevisiae Elp123 sub-complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.2 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : S.cerevisiae Elp123 sub-complex
Entire | Name: S.cerevisiae Elp123 sub-complex |
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Components |
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-Supramolecule #1: S.cerevisiae Elp123 sub-complex
Supramolecule | Name: S.cerevisiae Elp123 sub-complex / type: complex / ID: 1 / Parent: 0 Details: Composed of two copies of its three subunits: Elp1, Elp2 and Elp3. |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: BS1173 |
Molecular weight | Theoretical: 621 KDa |
-Experimental details
-Structure determination
Method | negative staining |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.035 mg/mL | ||||||||||||
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Buffer | pH: 7.6 Component:
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Staining | Type: NEGATIVE / Material: Uranyl Acetate Details: 3.5 ul aliquots of freshly purified Elp123 complex were applied to glow discharged carbon copper-collodion (Sigma) grids for 2 min and stained with a 1% uranyl acetate solution (w/v) | ||||||||||||
Grid | Model: Plano D-35578 / Material: COPPER / Mesh: 400 / Pretreatment - Type: GLOW DISCHARGE / Details: 0.45mBar, 20mA |
-Electron microscopy
Microscope | FEI TECNAI 12 |
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Image recording | Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number grids imaged: 1 / Number real images: 216 / Average electron dose: 16.0 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: LAB6 |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 6.3 mm / Nominal magnification: 49000 |