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Yorodumi- EMDB-30108: Cryo-Em structure of eukaryotic pre-60S ribosome subunit from Sac... -
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-Basic information
Entry | Database: EMDB / ID: EMD-30108 | ||||||||||||
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Title | Cryo-Em structure of eukaryotic pre-60S ribosome subunit from Saccharomyces cerevisiae rpf2 delta 255-344 strain, C4 state. | ||||||||||||
Map data | Cryo-EM structure of pre-60S ribosomal subunit from Saccharomyces cerevisiae rpf2 delta 255-344 strain, C4 state | ||||||||||||
Sample |
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Keywords | pre-60s / Rpf2 / RIBOSOME | ||||||||||||
Function / homology | Function and homology information protein-RNA complex remodeling / regulation of ribosomal subunit export from nucleus / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / nuclear exosome (RNase complex) / Hydrolases / PeBoW complex / 7S RNA binding / rRNA primary transcript binding / positive regulation of ATP-dependent activity / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...protein-RNA complex remodeling / regulation of ribosomal subunit export from nucleus / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / nuclear exosome (RNase complex) / Hydrolases / PeBoW complex / 7S RNA binding / rRNA primary transcript binding / positive regulation of ATP-dependent activity / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / 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) / proteasome binding / Major pathway of rRNA processing in the nucleolus and cytosol / 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 / nuclear-transcribed mRNA catabolic process / ATPase activator activity / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / regulation of translational fidelity / protein-RNA complex assembly / ribonucleoprotein complex binding / ribosomal subunit export from nucleus / 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) / maturation of LSU-rRNA / Neutrophil degranulation / translation initiation factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome complex / nuclear periphery / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / cytosolic ribosome assembly / small-subunit processome / macroautophagy / maintenance of translational fidelity / protein catabolic process / rRNA processing / metallopeptidase activity / protein transport / ribosome biogenesis / viral capsid / protein-macromolecule adaptor activity / ATPase binding / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / nucleic acid binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / GTPase activity / mRNA binding / host cell nucleus / GTP binding / nucleolus / proteolysis / RNA binding / zinc ion binding / nucleoplasm / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Saccharomyces cerevisiae S288C (yeast) / Saccharomyces cerevisiae S288c (yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||||||||
Authors | Li Y / Micic J | ||||||||||||
Funding support | China, 3 items
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Citation | Journal: Nat Commun / Year: 2020 Title: Coupling of 5S RNP rotation with maturation of functional centers during large ribosomal subunit assembly. Authors: Jelena Micic / Yu Li / Shan Wu / Daniel Wilson / Beril Tutuncuoglu / Ning Gao / John L Woolford / Abstract: The protein composition and structure of assembling 60S ribosomal subunits undergo numerous changes as pre-ribosomes transition from the nucleolus to the nucleoplasm. This includes stable anchoring ...The protein composition and structure of assembling 60S ribosomal subunits undergo numerous changes as pre-ribosomes transition from the nucleolus to the nucleoplasm. This includes stable anchoring of the Rpf2 subcomplex containing 5S rRNA, rpL5, rpL11, Rpf2 and Rrs1, which initially docks onto the flexible domain V of rRNA at earlier stages of assembly. In this work, we tested the function of the C-terminal domain (CTD) of Rpf2 during these anchoring steps, by truncating this extension and assaying effects on middle stages of subunit maturation. The rpf2Δ255-344 mutation affects proper folding of rRNA helices H68-70 during anchoring of the Rpf2 subcomplex. In addition, several assembly factors (AFs) are absent from pre-ribosomes or in altered conformations. Consequently, major remodeling events fail to occur: rotation of the 5S RNP, maturation of the peptidyl transferase center (PTC) and the nascent polypeptide exit tunnel (NPET), and export of assembling subunits to the cytoplasm. | ||||||||||||
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_30108.map.gz | 192.9 MB | EMDB map data format | |
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Header (meta data) | emd-30108-v30.xml emd-30108.xml | 74.8 KB 74.8 KB | Display Display | EMDB header |
Images | emd_30108.png | 69.5 KB | ||
Filedesc metadata | emd-30108.cif.gz | 17.1 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-30108 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-30108 | HTTPS FTP |
-Validation report
Summary document | emd_30108_validation.pdf.gz | 624.2 KB | Display | EMDB validaton report |
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Full document | emd_30108_full_validation.pdf.gz | 623.8 KB | Display | |
Data in XML | emd_30108_validation.xml.gz | 7.1 KB | Display | |
Data in CIF | emd_30108_validation.cif.gz | 8.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30108 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-30108 | HTTPS FTP |
-Related structure data
Related structure data | 6m62MC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_30108.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM structure of pre-60S ribosomal subunit from Saccharomyces cerevisiae rpf2 delta 255-344 strain, C4 state | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.057 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 : Eukaryotic pre-60S ribosomal subunits
+Supramolecule #1: Eukaryotic pre-60S ribosomal subunits
+Macromolecule #1: RDN25-1 rRNA
+Macromolecule #2: RDN58-1 rRNA
+Macromolecule #3: RDN5-2 rRNA
+Macromolecule #6: ITS2-1 miscRNA
+Macromolecule #4: Probable metalloprotease ARX1
+Macromolecule #5: rRNA-processing protein CGR1
+Macromolecule #7: 60S ribosomal protein L2-A
+Macromolecule #8: 60S ribosomal protein L3
+Macromolecule #9: 60S ribosomal protein L4-A
+Macromolecule #10: 60S ribosomal protein L5
+Macromolecule #11: 60S ribosomal protein L6-A
+Macromolecule #12: 60S ribosomal protein L7-A
+Macromolecule #13: 60S ribosomal protein L8-A
+Macromolecule #14: 60S ribosomal protein L9-A
+Macromolecule #15: Bud site selection protein 20
+Macromolecule #16: 60S ribosomal protein L11-A
+Macromolecule #17: Proteasome-interacting protein CIC1
+Macromolecule #18: 60S ribosomal protein L13-A
+Macromolecule #19: 60S ribosomal protein L14-A
+Macromolecule #20: 60S ribosomal protein L15-A
+Macromolecule #21: 60S ribosomal protein L16-A
+Macromolecule #22: 60S ribosomal protein L17-A
+Macromolecule #23: 60S ribosomal protein L18-A
+Macromolecule #24: 60S ribosomal protein L19-A
+Macromolecule #25: 60S ribosomal protein L20-A
+Macromolecule #26: 60S ribosomal protein L21-A
+Macromolecule #27: 60S ribosomal protein L22-A
+Macromolecule #28: 60S ribosomal protein L23-A
+Macromolecule #29: Ribosome assembly factor MRT4
+Macromolecule #30: 60S ribosomal protein L25
+Macromolecule #31: 60S ribosomal protein L26-A
+Macromolecule #32: 60S ribosomal protein L27-A
+Macromolecule #33: 60S ribosomal protein L28
+Macromolecule #34: Nucleolar GTP-binding protein 1
+Macromolecule #35: 60S ribosomal protein L30
+Macromolecule #36: 60S ribosomal protein L31-A
+Macromolecule #37: 60S ribosomal protein L32
+Macromolecule #38: 60S ribosomal protein L33-A
+Macromolecule #39: 60S ribosomal protein L34-A
+Macromolecule #40: 60S ribosomal protein L35-A
+Macromolecule #41: 60S ribosomal protein L36-A
+Macromolecule #42: 60S ribosomal protein L37-A
+Macromolecule #43: 60S ribosomal protein L38
+Macromolecule #44: 60S ribosomal protein L39
+Macromolecule #45: Nucleolar GTP-binding protein 2
+Macromolecule #46: Pescadillo homolog
+Macromolecule #47: Ribosome biogenesis protein 15
+Macromolecule #48: 60S ribosomal protein L43-A
+Macromolecule #49: Ribosome biogenesis protein NOP53
+Macromolecule #50: Ribosome biogenesis protein NSA2
+Macromolecule #51: Nuclear GTP-binding protein NUG1
+Macromolecule #52: Ribosome biogenesis protein RLP7
+Macromolecule #53: Ribosome biogenesis protein RLP24
+Macromolecule #54: Ribosome biogenesis protein RPF2
+Macromolecule #55: Regulator of ribosome biosynthesis
+Macromolecule #56: Ribosome assembly protein 4
+Macromolecule #57: Eukaryotic translation initiation factor 6
+Macromolecule #58: UPF0642 protein YBL028C
+Macromolecule #59: ZINC ION
+Macromolecule #60: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #61: 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) / Average electron dose: 1.9 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: OTHER |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: OTHER |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: EMDB MAP |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 53177 |
Initial angle assignment | Type: ANGULAR RECONSTITUTION |
Final angle assignment | Type: ANGULAR RECONSTITUTION |