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Yorodumi- EMDB-16541: Yeast cytoplasmic pre-40S ribosome biogenesis complex with deubiq... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-16541 | ||||||||||||||||||
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Title | Yeast cytoplasmic pre-40S ribosome biogenesis complex with deubiquitinating enzyme Otu2 | ||||||||||||||||||
Map data | Cryo-EM map of Otu2-bound cytoplasmic pre-40S ribosome biogenesis complex | ||||||||||||||||||
Sample |
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Keywords | Ribosome / Translation / Ubiquitin / Deubiquitinating enzyme / Biogenesis / Complex | ||||||||||||||||||
Function / homology | Function and homology information : / 18S rRNA (adenine(1779)-N(6)/adenine(1780)-N(6))-dimethyltransferase activity / intracellular organelle / negative regulation of glucose mediated signaling pathway / negative regulation of translational frameshifting / Negative regulators of DDX58/IFIH1 signaling / RMTs methylate histone arginines / positive regulation of translational fidelity / Protein methylation / mTORC1-mediated signalling ...: / 18S rRNA (adenine(1779)-N(6)/adenine(1780)-N(6))-dimethyltransferase activity / intracellular organelle / negative regulation of glucose mediated signaling pathway / negative regulation of translational frameshifting / Negative regulators of DDX58/IFIH1 signaling / RMTs methylate histone arginines / positive regulation of translational fidelity / Protein methylation / mTORC1-mediated signalling / Protein hydroxylation / ribosome-associated ubiquitin-dependent protein catabolic process / GDP-dissociation inhibitor activity / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / nonfunctional rRNA decay / preribosome, small subunit precursor / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / Ribosomal scanning and start codon recognition / Major pathway of rRNA processing in the nucleolus and cytosol / mRNA destabilization / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / 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 / regulation of amino acid metabolic process / L13a-mediated translational silencing of Ceruloplasmin expression / protein deubiquitination / 90S preribosome / G-protein alpha-subunit binding / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / Ub-specific processing proteases / ribosomal subunit export from nucleus / ribosomal small subunit export from nucleus / translation regulator activity / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / DNA-(apurinic or apyrimidinic site) endonuclease activity / RNA endonuclease activity / rescue of stalled ribosome / Transferases; Transferring one-carbon groups; Methyltransferases / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / translational initiation / protein kinase C binding / maintenance of translational fidelity / cytoplasmic stress granule / rRNA processing / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosome biogenesis / ribosome binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / cytoplasmic translation / cysteine-type deubiquitinase activity / negative regulation of translation / non-specific serine/threonine protein kinase / rRNA binding / ribosome / structural constituent of ribosome / positive regulation of protein phosphorylation / protein phosphorylation / G protein-coupled receptor signaling pathway / translation / negative regulation of gene expression / protein serine/threonine kinase activity / mRNA binding / nucleolus / mitochondrion / RNA binding / zinc ion binding / nucleoplasm / ATP binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||||||||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) / Saccharomyces cerevisiae W303 (yeast) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | ||||||||||||||||||
Authors | Ikeuchi K / Buschauer R / Cheng J / Berninghausen O / Becker T / Beckmann R | ||||||||||||||||||
Funding support | Germany, Japan, 5 items
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Citation | Journal: Nat Commun / Year: 2023 Title: Molecular basis for recognition and deubiquitination of 40S ribosomes by Otu2. Authors: Ken Ikeuchi / Nives Ivic / Robert Buschauer / Jingdong Cheng / Thomas Fröhlich / Yoshitaka Matsuo / Otto Berninghausen / Toshifumi Inada / Thomas Becker / Roland Beckmann / Abstract: In actively translating 80S ribosomes the ribosomal protein eS7 of the 40S subunit is monoubiquitinated by the E3 ligase Not4 and deubiquitinated by Otu2 upon ribosomal subunit recycling. Despite its ...In actively translating 80S ribosomes the ribosomal protein eS7 of the 40S subunit is monoubiquitinated by the E3 ligase Not4 and deubiquitinated by Otu2 upon ribosomal subunit recycling. Despite its importance for translation efficiency the exact role and structural basis for this translational reset is poorly understood. Here, structural analysis by cryo-electron microscopy of native and reconstituted Otu2-bound ribosomal complexes reveals that Otu2 engages 40S subunits mainly between ribosome recycling and initiation stages. Otu2 binds to several sites on the intersubunit surface of the 40S that are not occupied by any other 40S-binding factors. This binding mode explains the discrimination against 80S ribosomes via the largely helical N-terminal domain of Otu2 as well as the specificity for mono-ubiquitinated eS7 on 40S. Collectively, this study reveals mechanistic insights into the Otu2-driven deubiquitination steps for translational reset during ribosome recycling/(re)initiation. | ||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_16541.map.gz | 13.8 MB | EMDB map data format | |
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Header (meta data) | emd-16541-v30.xml emd-16541.xml | 59 KB 59 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_16541_fsc.xml | 14 KB | Display | FSC data file |
Images | emd_16541.png | 134.6 KB | ||
Filedesc metadata | emd-16541.cif.gz | 13.9 KB | ||
Others | emd_16541_half_map_1.map.gz emd_16541_half_map_2.map.gz | 262.2 MB 262.1 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-16541 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-16541 | HTTPS FTP |
-Validation report
Summary document | emd_16541_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_16541_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_16541_validation.xml.gz | 23.3 KB | Display | |
Data in CIF | emd_16541_validation.cif.gz | 30.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-16541 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-16541 | HTTPS FTP |
-Related structure data
Related structure data | 8cbjMC 8c83C 8cahC 8casC C: citing same article (ref.) M: atomic model generated by this map |
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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_16541.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM map of Otu2-bound cytoplasmic pre-40S ribosome biogenesis complex | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.045 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_16541_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_16541_half_map_2.map | ||||||||||||
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Annotation | Half-map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Yeast cytoplasmic pre-40S ribosome biogenesis complex with deubiq...
+Supramolecule #1: Yeast cytoplasmic pre-40S ribosome biogenesis complex with deubiq...
+Macromolecule #1: 40S ribosomal protein S27-A
+Macromolecule #2: 40S ribosomal protein S28-A
+Macromolecule #3: RPS29A isoform 1
+Macromolecule #4: 40S ribosomal protein S30-A
+Macromolecule #5: Guanine nucleotide-binding protein subunit beta-like protein
+Macromolecule #6: Pre-rRNA-processing protein PNO1
+Macromolecule #7: ENP1 isoform 1
+Macromolecule #8: LTV1 isoform 1
+Macromolecule #9: TSR1 isoform 1
+Macromolecule #10: non-specific serine/threonine protein kinase
+Macromolecule #11: 40S ribosomal protein S0-A
+Macromolecule #12: 40S ribosomal protein S1-A
+Macromolecule #13: 40S ribosomal protein S2
+Macromolecule #14: 40S ribosomal protein S3
+Macromolecule #15: 40S ribosomal protein S4-B
+Macromolecule #16: 40S ribosomal protein S5
+Macromolecule #17: 40S ribosomal protein S6-B
+Macromolecule #18: 40S ribosomal protein S7-A
+Macromolecule #19: 40S ribosomal protein S8-A
+Macromolecule #20: 40S ribosomal protein S9-A
+Macromolecule #21: 40S ribosomal protein S11-A
+Macromolecule #22: 40S ribosomal protein S12
+Macromolecule #23: 40S ribosomal protein S13
+Macromolecule #24: 40S ribosomal protein S14-A
+Macromolecule #25: RPS15 isoform 1
+Macromolecule #26: 40S ribosomal protein S16-A
+Macromolecule #27: 40S ribosomal protein S17-A
+Macromolecule #28: 40S ribosomal protein S18-A
+Macromolecule #29: 40S ribosomal protein S19-A
+Macromolecule #30: 40S ribosomal protein S20
+Macromolecule #31: 40S ribosomal protein S21-A
+Macromolecule #32: 40S ribosomal protein S22-A
+Macromolecule #33: 40S ribosomal protein S23-A
+Macromolecule #34: 40S ribosomal protein S24-A
+Macromolecule #35: 40S ribosomal protein S25-A
+Macromolecule #36: 20S-pre-rRNA D-site endonuclease NOB1
+Macromolecule #37: rRNA adenine N(6)-methyltransferase
+Macromolecule #39: OTU domain-containing protein 2
+Macromolecule #38: 20S pre-ribosomal RNA
+Macromolecule #40: ZINC ION
+Macromolecule #41: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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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) / Detector mode: COUNTING / Average electron dose: 46.4 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: 3.0 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |