+ Open data
Open data
- Basic information
Basic information
| Entry | Database: PDB / ID: 8fle | |||||||||
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| Title | Human nuclear pre-60S ribosomal subunit (State L2) | |||||||||
|  Components | 
 | |||||||||
|  Keywords | RIBOSOME / Pre-60S ribosomal subunit / Assembly intermediate / Nucleoprotein complex | |||||||||
| Function / homology |  Function and homology information protein localization to nucleoplasm / negative regulation of transcription of nucleolar large rRNA by RNA polymerase I / negative regulation of RNA polymerase II regulatory region sequence-specific DNA binding / regulation of RIG-I signaling pathway / basal RNA polymerase II transcription machinery binding / dendrite extension / preribosome binding / lamin filament / regulation of fatty acid biosynthetic process / regulation of megakaryocyte differentiation ...protein localization to nucleoplasm / negative regulation of transcription of nucleolar large rRNA by RNA polymerase I / negative regulation of RNA polymerase II regulatory region sequence-specific DNA binding / regulation of RIG-I signaling pathway / basal RNA polymerase II transcription machinery binding / dendrite extension / preribosome binding / lamin filament / regulation of fatty acid biosynthetic process / regulation of megakaryocyte differentiation / miRNA-mediated post-transcriptional gene silencing / negative regulation of G2/M transition of mitotic cell cycle / PeBoW complex / miRNA-mediated gene silencing by inhibition of translation / translation at presynapse / embryonic brain development / eukaryotic 80S initiation complex / negative regulation of protein neddylation / regulation of translation involved in cellular response to UV / axial mesoderm development / negative regulation of formation of translation preinitiation complex / regulation of G1 to G0 transition / positive regulation of protein K63-linked deubiquitination / ribosomal protein import into nucleus / blastocyst formation / protein localization to nucleolus / protein-DNA complex disassembly / 90S preribosome assembly / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / GAIT complex / positive regulation of DNA damage response, signal transduction by p53 class mediator / TORC2 complex binding / alpha-beta T cell differentiation / G1 to G0 transition / regulation of glycolytic process / middle ear morphogenesis / regulation of reactive oxygen species metabolic process / negative regulation of cell-cell adhesion / regulation of aerobic respiration / cytoplasmic side of rough endoplasmic reticulum membrane / maturation of 5.8S rRNA / negative regulation of ubiquitin protein ligase activity / homeostatic process / negative regulation of DNA replication / rRNA metabolic process / macrophage chemotaxis / positive regulation of dendritic spine development / lung morphogenesis / negative regulation of signal transduction by p53 class mediator / mitotic G2 DNA damage checkpoint signaling / positive regulation of natural killer cell proliferation / ribosomal large subunit binding / regulation of protein phosphorylation / Protein hydroxylation / preribosome, large subunit precursor / Peptide chain elongation / nuclear-transcribed mRNA catabolic process / rRNA transcription / Selenocysteine synthesis / Formation of a pool of free 40S subunits / Eukaryotic Translation Termination / ubiquitin ligase inhibitor activity / cellular response to actinomycin D / blastocyst development / Response of EIF2AK4 (GCN2) to amino acid deficiency / SRP-dependent cotranslational protein targeting to membrane / negative regulation of ubiquitin-dependent protein catabolic process / positive regulation of signal transduction by p53 class mediator / Viral mRNA Translation / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / protein localization to nucleus / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / positive regulation of protein binding / Major pathway of rRNA processing in the nucleolus and cytosol / ribosomal subunit export from nucleus / ribonucleoprotein complex binding / protein targeting / negative regulation of protein-containing complex assembly / protein-RNA complex assembly / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / maturation of LSU-rRNA / rough endoplasmic reticulum / translation regulator activity / MDM2/MDM4 family protein binding / translation initiation factor activity / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / negative regulation of protein ubiquitination / embryo implantation / cytosolic ribosome / negative regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction / cellular response to interleukin-4 / ossification / negative regulation of cell migration / assembly of large subunit precursor of preribosome / positive regulation of translation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of signal transduction by p53 class mediator / ribosomal large subunit biogenesis Similarity search - Function | |||||||||
| Biological species |  Homo sapiens (human) | |||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.48 Å | |||||||||
|  Authors | Vanden Broeck, A. / Klinge, S. | |||||||||
| Funding support | European Union,  United States, 2items 
 | |||||||||
|  Citation |  Journal: Science / Year: 2023 Title: Principles of human pre-60 biogenesis. Authors: Arnaud Vanden Broeck / Sebastian Klinge /  Abstract: During the early stages of human large ribosomal subunit (60) biogenesis, an ensemble of assembly factors establishes and fine-tunes the essential RNA functional centers of pre-60 particles by an ...During the early stages of human large ribosomal subunit (60) biogenesis, an ensemble of assembly factors establishes and fine-tunes the essential RNA functional centers of pre-60 particles by an unknown mechanism. Here, we report a series of cryo-electron microscopy structures of human nucleolar and nuclear pre-60 assembly intermediates at resolutions of 2.5 to 3.2 angstroms. These structures show how protein interaction hubs tether assembly factor complexes to nucleolar particles and how guanosine triphosphatases and adenosine triphosphatase couple irreversible nucleotide hydrolysis steps to the installation of functional centers. Nuclear stages highlight how a conserved RNA-processing complex, the rixosome, couples large-scale RNA conformational changes with pre-ribosomal RNA processing by the RNA degradation machinery. Our ensemble of human pre-60 particles provides a rich foundation with which to elucidate the molecular principles of ribosome formation. | |||||||||
| History | 
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- Structure visualization
Structure visualization
| Structure viewer | Molecule:  Molmil  Jmol/JSmol | 
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- Downloads & links
Downloads & links
- Download
Download
| PDBx/mmCIF format |  8fle.cif.gz | 3.2 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb8fle.ent.gz | Display |  PDB format | |
| PDBx/mmJSON format |  8fle.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  8fle_validation.pdf.gz | 1.9 MB | Display |  wwPDB validaton report | 
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| Full document |  8fle_full_validation.pdf.gz | 2 MB | Display | |
| Data in XML |  8fle_validation.xml.gz | 262.4 KB | Display | |
| Data in CIF |  8fle_validation.cif.gz | 437 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/fl/8fle  ftp://data.pdbj.org/pub/pdb/validation_reports/fl/8fle | HTTPS FTP | 
-Related structure data
| Related structure data |  29276MC  8fkpC  8fkqC  8fkrC  8fksC  8fktC  8fkuC  8fkvC  8fkwC  8fkxC  8fkyC  8fkzC  8fl0C  8fl2C  8fl3C  8fl4C  8fl6C  8fl7C  8fl9C  8flaC  8flbC  8flcC  8fldC  8flfC C: citing same article ( M: map data used to model this data | 
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| Similar structure data | Similarity search - Function & homology  F&H Search | 
- Links
Links
- Assembly
Assembly
| Deposited unit |  
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| 1 | 
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- Components
Components
+60S ribosomal protein  ... , 40 types, 40 molecules BEL5L6L7L8L9LALBLCLDLELFLGLHLILJLKLLLMLNLOLPLQLRLSLTLULVLWLX...                              
-RNA chain , 3 types, 3 molecules L1L3L4  
| #2: RNA chain | Mass: 50463.840 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: GenBank: 555853 | 
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| #3: RNA chain | Mass: 1641203.750 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: GenBank: 86475748 | 
| #4: RNA chain | Mass: 38998.078 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: GenBank: 23898 | 
-Protein , 9 types, 9 molecules NKNLNPSHSKSMSQSRSV        
| #36: Protein | Mass: 15268.361 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: Q9BRT6 | 
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| #37: Protein | Mass: 54498.402 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: Q9NZM5 | 
| #38: Protein | Mass: 15230.225 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: O00488 | 
| #46: Protein | Mass: 34285.309 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: Q9BYG3 | 
| #48: Protein | Mass: 26620.010 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: P56537 | 
| #49: Protein | Mass: 68114.727 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: O00541 | 
| #50: Protein | Mass: 27602.535 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: Q9UKD2 | 
| #51: Protein | Mass: 74107.820 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: Q9BZE4 | 
| #52: Protein | Mass: 19666.258 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / Cell line: HEK293F / References: UniProt: Q9UHA3 | 
-Non-polymers , 2 types, 93 molecules 


| #53: Chemical | ChemComp-MG / #54: Chemical | ChemComp-ZN / | 
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-Details
| Has ligand of interest | N | 
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-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY | 
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction | 
- Sample preparation
Sample preparation
| Component | Name: Human nuclear pre-60S ribosomal subunit (State L2) / Type: RIBOSOME / Entity ID: #1-#52 / Source: NATURAL | 
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| Molecular weight | Experimental value: NO | 
| Source (natural) | Organism:  Homo sapiens (human) / Strain: HEK293F | 
| Buffer solution | pH: 7.6 | 
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | 
| Specimen support | Grid material: GOLD / Grid mesh size: 400 divisions/in. / Grid type: Quantifoil R3.5/1 | 
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 95 % / Chamber temperature: 283 K Details: Four applications with manual blotting before last blotting with the Vitrobot. | 
- Electron microscopy imaging
Electron microscopy imaging
| Experimental equipment |  Model: Titan Krios / Image courtesy: FEI Company | 
|---|---|
| Microscopy | Model: FEI TITAN KRIOS | 
| Electron gun | Electron source:  FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM | 
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 64000 X / Nominal defocus max: 2500 nm / Nominal defocus min: 500 nm | 
| Image recording | Average exposure time: 2 sec. / Electron dose: 60 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 4 / Num. of real images: 172699 | 
| EM imaging optics | Energyfilter slit width: 20 eV | 
- Processing
Processing
| EM software | 
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 15679142 | ||||||||||||||||||||||||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 2.48 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 123749 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | Space: REAL | 
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