+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-15423 | |||||||||
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Title | Yeast RQC complex in state F | |||||||||
Map data | Yeast RQC complex in state G | |||||||||
Sample |
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Keywords | ribosome-associated quality control / NEMF / Listerin / CAT tailing / TRANSLATION | |||||||||
Function / homology | Function and homology information alpha-aminoacyl-tRNA binding / positive regulation of cytoplasmic translational elongation through polyproline stretches / Hypusine synthesis from eIF5A-lysine / CAT tailing / translational frameshifting / positive regulation of translational termination / RQC complex / peptide biosynthetic process / positive regulation of translational elongation / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...alpha-aminoacyl-tRNA binding / positive regulation of cytoplasmic translational elongation through polyproline stretches / Hypusine synthesis from eIF5A-lysine / CAT tailing / translational frameshifting / positive regulation of translational termination / RQC complex / peptide biosynthetic process / positive regulation of translational elongation / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosome-associated ubiquitin-dependent protein catabolic process / maturation of 5.8S rRNA / hexon binding / pre-mRNA 5'-splice site binding / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / response to cycloheximide / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / ribosomal large subunit binding / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Formation of a pool of free 40S subunits / negative regulation of mRNA splicing, via spliceosome / preribosome, large subunit precursor / Antigen processing: Ubiquitination & Proteasome degradation / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / 90S preribosome / subtelomeric heterochromatin formation / regulation of translational fidelity / positive regulation of translational initiation / protein-RNA complex assembly / proteasomal protein catabolic process / translation elongation factor activity / ribosomal subunit export from nucleus / translational termination / maturation of LSU-rRNA / cytosolic ribosome / translation initiation factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rescue of stalled ribosome / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / cytosolic ribosome assembly / translational initiation / macroautophagy / maintenance of translational fidelity / RING-type E3 ubiquitin transferase / protein catabolic process / modification-dependent protein catabolic process / cytoplasmic stress granule / ubiquitin-protein transferase activity / protein tag activity / rRNA processing / ubiquitin protein ligase activity / ribosome biogenesis / viral capsid / ribosome binding / chromatin organization / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / ubiquitin-dependent protein catabolic process / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / chromosome, telomeric region / negative regulation of translation / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / translation / response to antibiotic / mRNA binding / ubiquitin protein ligase binding / host cell nucleus / nucleolus / perinuclear region of cytoplasm / mitochondrion / RNA binding / zinc ion binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.6 Å | |||||||||
Authors | Tesina P / Buschauer R / Beckmann R | |||||||||
Funding support | Germany, 1 items
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Citation | Journal: Mol Cell / Year: 2023 Title: Molecular basis of eIF5A-dependent CAT tailing in eukaryotic ribosome-associated quality control. Authors: Petr Tesina / Shuhei Ebine / Robert Buschauer / Matthias Thoms / Yoshitaka Matsuo / Toshifumi Inada / Roland Beckmann / Abstract: Ribosome-associated quality control (RQC) is a conserved process degrading potentially toxic truncated nascent peptides whose malfunction underlies neurodegeneration and proteostasis decline in aging. ...Ribosome-associated quality control (RQC) is a conserved process degrading potentially toxic truncated nascent peptides whose malfunction underlies neurodegeneration and proteostasis decline in aging. During RQC, dissociation of stalled ribosomes is followed by elongation of the nascent peptide with alanine and threonine residues, driven by Rqc2 independently of mRNA, the small ribosomal subunit and guanosine triphosphate (GTP)-hydrolyzing factors. The resulting CAT tails (carboxy-terminal tails) and ubiquitination by Ltn1 mark nascent peptides for proteasomal degradation. Here we present ten cryogenic electron microscopy (cryo-EM) structures, revealing the mechanistic basis of individual steps of the CAT tailing cycle covering initiation, decoding, peptidyl transfer, and tRNA translocation. We discovered eIF5A as a crucial eukaryotic RQC factor enabling peptidyl transfer. Moreover, we observed dynamic behavior of RQC factors and tRNAs allowing for processivity of the CAT tailing cycle without additional energy input. Together, these results elucidate key differences as well as common principles between CAT tailing and canonical translation. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_15423.map.gz | 20.4 MB | EMDB map data format | |
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Header (meta data) | emd-15423-v30.xml emd-15423.xml | 75 KB 75 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_15423_fsc.xml | 15.6 KB | Display | FSC data file |
Images | emd_15423.png | 60.2 KB | ||
Filedesc metadata | emd-15423.cif.gz | 15.6 KB | ||
Others | emd_15423_half_map_1.map.gz emd_15423_half_map_2.map.gz | 322.7 MB 322.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-15423 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-15423 | HTTPS FTP |
-Validation report
Summary document | emd_15423_validation.pdf.gz | 892.3 KB | Display | EMDB validaton report |
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Full document | emd_15423_full_validation.pdf.gz | 891.9 KB | Display | |
Data in XML | emd_15423_validation.xml.gz | 24.5 KB | Display | |
Data in CIF | emd_15423_validation.cif.gz | 31.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15423 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15423 | HTTPS FTP |
-Related structure data
Related structure data | 8agtMC 8aafC 8aguC 8agvC 8agwC 8agxC 8agzC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_15423.map.gz / Format: CCP4 / Size: 347.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Yeast RQC complex in state G | ||||||||||||||||||||||||||||||||||||
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: Yeast RQC complex in state G, half map A
File | emd_15423_half_map_1.map | ||||||||||||
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Annotation | Yeast RQC complex in state G, half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Yeast RQC complex in state G, half map B
File | emd_15423_half_map_2.map | ||||||||||||
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Annotation | Yeast RQC complex in state G, half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Yeast RQC complex in state F
+Supramolecule #1: Yeast RQC complex in state F
+Macromolecule #1: 60S ribosomal protein L15-A
+Macromolecule #2: 60S ribosomal protein L16-A
+Macromolecule #3: 60S ribosomal protein L17-A
+Macromolecule #4: 60S ribosomal protein L18-A
+Macromolecule #5: 60S ribosomal protein L19-A
+Macromolecule #6: 60S ribosomal protein L20-A
+Macromolecule #7: 60S ribosomal protein L21-A
+Macromolecule #8: 60S ribosomal protein L22-A
+Macromolecule #9: 60S ribosomal protein L23-A
+Macromolecule #10: 60S ribosomal protein L24-A
+Macromolecule #11: 60S ribosomal protein L25
+Macromolecule #12: 60S ribosomal protein L26-A
+Macromolecule #13: 60S ribosomal protein L27-A
+Macromolecule #14: 60S ribosomal protein L28
+Macromolecule #15: 60S ribosomal protein L29
+Macromolecule #16: 60S ribosomal protein L30
+Macromolecule #17: 60S ribosomal protein L31-A
+Macromolecule #18: 60S ribosomal protein L32
+Macromolecule #19: 60S ribosomal protein L33-A
+Macromolecule #20: 60S ribosomal protein L34-A
+Macromolecule #21: 60S ribosomal protein L35-A
+Macromolecule #22: 60S ribosomal protein L36-A
+Macromolecule #23: 60S ribosomal protein L37-A
+Macromolecule #24: 60S ribosomal protein L38
+Macromolecule #25: 60S ribosomal protein L39
+Macromolecule #26: Ubiquitin-60S ribosomal protein L40
+Macromolecule #27: 60S ribosomal protein L42-A
+Macromolecule #28: 60S ribosomal protein L43-A
+Macromolecule #29: 60S ribosomal protein L41-A
+Macromolecule #33: 60S ribosomal protein L2-A
+Macromolecule #34: 60S ribosomal protein L3
+Macromolecule #35: 60S ribosomal protein L4-A
+Macromolecule #36: 60S ribosomal protein L5
+Macromolecule #37: 60S ribosomal protein L6-B
+Macromolecule #38: 60S ribosomal protein L7-A
+Macromolecule #39: 60S ribosomal protein L8-A
+Macromolecule #40: 60S ribosomal protein L9-A
+Macromolecule #41: 60S ribosomal protein L10
+Macromolecule #42: 60S ribosomal protein L11-A
+Macromolecule #43: 60S ribosomal protein L13-A
+Macromolecule #44: 60S ribosomal protein L14-A
+Macromolecule #45: Ribosome quality control complex subunit 2
+Macromolecule #46: E3 ubiquitin-protein ligase listerin
+Macromolecule #47: Eukaryotic translation initiation factor 6
+Macromolecule #48: 60S ribosomal protein L1-A
+Macromolecule #51: 60S ribosomal protein L12-A
+Macromolecule #52: 60S acidic ribosomal protein P0
+Macromolecule #53: CAT tailed nascent peptide
+Macromolecule #54: Eukaryotic translation initiation factor 5A-1
+Macromolecule #30: 25S rRNA
+Macromolecule #31: 5S rRNA
+Macromolecule #32: 5.8S rRNA
+Macromolecule #49: A-site Ala aminoacyl-tRNA
+Macromolecule #50: P-site petidyl-tRNA
+Macromolecule #55: MAGNESIUM ION
+Macromolecule #56: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 6.8 |
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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: 46.0 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: 4.0 µm / Nominal defocus min: 0.4 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |