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- EMDB-2179: Electron cryo-microscopy of C. thermophilum RAC bound to the 80S ... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2179 | |||||||||
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Title | Electron cryo-microscopy of C. thermophilum RAC bound to the 80S ribosome | |||||||||
![]() | Reconstruction of the reconstituted complex of C. thermophilum RAC and C. thermophilum 80S ribosomes (Low-pass filtered between 18-20 A) | |||||||||
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![]() | ribosome-associated complex / RAC / ribosome / 80S / chaperone / co-translational folding | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 9.6 Å | |||||||||
![]() | Leidig C / Bange G / Kopp J / Amlacher S / Aravind A / Wickles S / Witte G / Hurt E / Beckmann R / Sinning I | |||||||||
![]() | ![]() Title: Structural characterization of a eukaryotic chaperone--the ribosome-associated complex. Authors: Christoph Leidig / Gert Bange / Jürgen Kopp / Stefan Amlacher / Ajay Aravind / Stephan Wickles / Gregor Witte / Ed Hurt / Roland Beckmann / Irmgard Sinning / ![]() Abstract: Ribosome-associated chaperones act in early folding events during protein synthesis. Structural information is available for prokaryotic chaperones (such as trigger factor), but structural ...Ribosome-associated chaperones act in early folding events during protein synthesis. Structural information is available for prokaryotic chaperones (such as trigger factor), but structural understanding of these processes in eukaryotes lags far behind. Here we present structural analyses of the eukaryotic ribosome-associated complex (RAC) from Saccharomyces cerevisiae and Chaetomium thermophilum, consisting of heat-shock protein 70 (Hsp70) Ssz1 and the Hsp40 Zuo1. RAC is an elongated complex that crouches over the ribosomal tunnel exit and seems to be stabilized in a distinct conformation by expansion segment ES27. A unique α-helical domain in Zuo1 mediates ribosome interaction of RAC near the ribosomal proteins L22e and L31e and ribosomal RNA helix H59. The crystal structure of the Ssz1 ATPase domain bound to ATP-Mg²⁺ explains its catalytic inactivity and suggests that Ssz1 may act before the RAC-associated chaperone Ssb. Our study offers insights into the interplay between RAC, the ER membrane-integrated Hsp40-type protein ERj1 and the signal-recognition particle. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 3.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 9.8 KB 9.8 KB | Display Display | ![]() |
Images | ![]() | 117 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Reconstruction of the reconstituted complex of C. thermophilum RAC and C. thermophilum 80S ribosomes (Low-pass filtered between 18-20 A) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.1 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Ribosome-associated complex (RAC) bound to 80S ribosome
Entire | Name: Ribosome-associated complex (RAC) bound to 80S ribosome |
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Components |
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-Supramolecule #1000: Ribosome-associated complex (RAC) bound to 80S ribosome
Supramolecule | Name: Ribosome-associated complex (RAC) bound to 80S ribosome type: sample / ID: 1000 Details: Sample was reconstituted from the purified components Oligomeric state: One RAC binds to one ribosome / Number unique components: 2 |
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Molecular weight | Theoretical: 3.2 MDa |
-Supramolecule #1: 80S ribosome
Supramolecule | Name: 80S ribosome / type: complex / ID: 1 / Recombinant expression: No / Ribosome-details: ribosome-eukaryote: ALL |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 3.1 MDa |
-Macromolecule #1: Ribosome-associated complex
Macromolecule | Name: Ribosome-associated complex / type: protein_or_peptide / ID: 1 / Name.synonym: RAC / Recombinant expression: No |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 110 KDa |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7 Details: 20 mM Tris-HCl, 100 mM KOAc, 100 mM sucrose, 7.5 mM Mg(OAc)2, 2 mM DTT |
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Grid | Details: carbon supported |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Instrument: FEI VITROBOT MARK III / Method: Blot for 3.5 s before plunging |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Details | Final magnification of the object on the CCD image is 148721 |
Date | Jul 6, 2011 |
Image recording | Category: CCD / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Number real images: 6954 / Average electron dose: 20 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.7 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 75000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Details | Particles selected by signature. Refinement with spider. |
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CTF correction | Details: Particle groups with similar defocus |
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 9.6 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: SPIDER Details: Sorting for ribosome conformation and ligand presence was performed Number images used: 26038 |