+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10842 | |||||||||
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Title | pre-60S State NE2 (TAP-Flag-Nop53) | |||||||||
Map data | State NE2: pre-60S ribosomal subunit purified via Nop53 Local resolution filtered map | |||||||||
Sample |
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Keywords | pre-60S / Biogenesis / LSU / Large subunit / ribosome assembly / RIBOSOME | |||||||||
Function / homology | Function and homology information 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) / 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) / 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) / maturation of 5.8S rRNA / proteasome binding / Major pathway of rRNA processing in the nucleolus and cytosol ...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) / 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) / 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) / maturation of 5.8S 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 / maturation of LSU-rRNA / translation initiation factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome complex / 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 / protein transport / ribosome biogenesis / ATPase binding / protein-macromolecule adaptor activity / large ribosomal subunit rRNA binding / 5S 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 / ribonucleoprotein complex / GTPase activity / mRNA binding / nucleolus / GTP binding / RNA binding / zinc ion binding / nucleoplasm / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||
Authors | Kater L / Beckmann R | |||||||||
Citation | Journal: Mol Cell / Year: 2020 Title: Construction of the Central Protuberance and L1 Stalk during 60S Subunit Biogenesis. Authors: Lukas Kater / Valentin Mitterer / Matthias Thoms / Jingdong Cheng / Otto Berninghausen / Roland Beckmann / Ed Hurt / Abstract: Ribosome assembly is driven by numerous assembly factors, including the Rix1 complex and the AAA ATPase Rea1. These two assembly factors catalyze 60S maturation at two distinct states, triggering ...Ribosome assembly is driven by numerous assembly factors, including the Rix1 complex and the AAA ATPase Rea1. These two assembly factors catalyze 60S maturation at two distinct states, triggering poorly understood large-scale structural transitions that we analyzed by cryo-electron microscopy. Two nuclear pre-60S intermediates were discovered that represent previously unknown states after Rea1-mediated removal of the Ytm1-Erb1 complex and reveal how the L1 stalk develops from a pre-mature nucleolar to a mature-like nucleoplasmic state. A later pre-60S intermediate shows how the central protuberance arises, assisted by the nearby Rix1-Rea1 machinery, which was solved in its pre-ribosomal context to molecular resolution. This revealed a Rix1-Ipi3 tetramer anchored to the pre-60S via Ipi1, strategically positioned to monitor this decisive remodeling. These results are consistent with a general underlying principle that temporarily stabilized immature RNA domains are successively remodeled by assembly factors, thereby ensuring failsafe assembly progression. | |||||||||
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_10842.map.gz | 126.5 MB | EMDB map data format | |
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Header (meta data) | emd-10842-v30.xml emd-10842.xml | 70.5 KB 70.5 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_10842_fsc.xml | 13.7 KB | Display | FSC data file |
Images | emd_10842.png | 120.9 KB | ||
Filedesc metadata | emd-10842.cif.gz | 15.5 KB | ||
Others | emd_10842_half_map_1.map.gz emd_10842_half_map_2.map.gz | 170.9 MB 170.9 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10842 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10842 | HTTPS FTP |
-Validation report
Summary document | emd_10842_validation.pdf.gz | 882.8 KB | Display | EMDB validaton report |
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Full document | emd_10842_full_validation.pdf.gz | 882.3 KB | Display | |
Data in XML | emd_10842_validation.xml.gz | 21.2 KB | Display | |
Data in CIF | emd_10842_validation.cif.gz | 27.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10842 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10842 | HTTPS FTP |
-Related structure data
Related structure data | 6ylyMC 6yleC 6ylfC 6ylgC 6ylhC 6ylxC 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_10842.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | State NE2: pre-60S ribosomal subunit purified via Nop53 Local resolution filtered map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.084 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: Relion half map (unfiltered, unmasked)
File | emd_10842_half_map_1.map | ||||||||||||
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Annotation | Relion half map (unfiltered, unmasked) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Relion half map (unfiltered, unmasked)
File | emd_10842_half_map_2.map | ||||||||||||
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Annotation | Relion half map (unfiltered, unmasked) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : pre-60S assembly state NE2
+Supramolecule #1: pre-60S assembly state NE2
+Macromolecule #1: 60S ribosomal protein L2-A
+Macromolecule #2: 60S ribosomal protein L28
+Macromolecule #3: 60S ribosomal protein L3
+Macromolecule #4: Nucleolar GTP-binding protein 1
+Macromolecule #5: 60S ribosomal protein L4-A
+Macromolecule #6: 60S ribosomal protein L30
+Macromolecule #7: 60S ribosomal protein L31-A
+Macromolecule #8: 60S ribosomal protein L6-A
+Macromolecule #9: 60S ribosomal protein L32
+Macromolecule #10: 60S ribosomal protein L7-A
+Macromolecule #11: 60S ribosomal protein L33-A
+Macromolecule #12: 60S ribosomal protein L8-A
+Macromolecule #13: 60S ribosomal protein L34-A
+Macromolecule #14: Bud site selection protein 20
+Macromolecule #15: 60S ribosomal protein L9-A
+Macromolecule #16: 60S ribosomal protein L35-A
+Macromolecule #17: 60S ribosomal protein L36-A
+Macromolecule #18: 60S ribosomal protein L37-A
+Macromolecule #19: 60S ribosomal protein L38
+Macromolecule #20: 60S ribosomal protein L13-A
+Macromolecule #21: 60S ribosomal protein L39
+Macromolecule #22: 60S ribosomal protein L14-A
+Macromolecule #23: 60S ribosomal protein L43-A
+Macromolecule #24: 60S ribosomal protein L15-A
+Macromolecule #25: Ribosome biogenesis protein RLP24
+Macromolecule #26: 60S ribosomal protein L16-A
+Macromolecule #27: 60S ribosomal protein L17-A
+Macromolecule #28: 60S ribosomal protein L18-A
+Macromolecule #29: 60S ribosomal protein L19-A
+Macromolecule #30: Ribosome biogenesis protein NSA2
+Macromolecule #31: 60S ribosomal protein L20-A
+Macromolecule #32: 60S ribosomal protein L21-A
+Macromolecule #33: 60S ribosomal protein L22-A
+Macromolecule #34: 60S ribosomal protein L23-A
+Macromolecule #35: Ribosome assembly factor MRT4
+Macromolecule #36: 60S ribosomal protein L25
+Macromolecule #37: 60S ribosomal protein L26-A
+Macromolecule #38: Eukaryotic translation initiation factor 6
+Macromolecule #39: 60S ribosomal protein L27-A
+Macromolecule #40: UPF0642 protein YBL028C
+Macromolecule #43: Proteasome-interacting protein CIC1
+Macromolecule #44: Pescadillo homolog
+Macromolecule #45: Ribosome biogenesis protein 15
+Macromolecule #46: Ribosome biogenesis protein NOP53
+Macromolecule #47: Ribosome biogenesis protein RLP7
+Macromolecule #49: Nuclear GTP-binding protein NUG1
+Macromolecule #41: 25S rRNA
+Macromolecule #42: 5.8S rRNA
+Macromolecule #48: ITS2
+Macromolecule #50: ZINC 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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Grid | Model: Quantifoil R3/3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Digitization - Frames/image: 1-10 / Number grids imaged: 2 / Average electron dose: 24.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |