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Yorodumi- PDB-6em3: State A architectural model (Nsa1-TAP Flag-Ytm1) - Visualizing th... -
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Basic information
| Entry | Database: PDB / ID: 6em3 | ||||||
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| Title | State A architectural model (Nsa1-TAP Flag-Ytm1) - Visualizing the assembly pathway of nucleolar pre-60S ribosomes | ||||||
Components |
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Keywords | RIBOSOME / Large Subunit Biogenesis Nucleolus | ||||||
| Function / homology | Function and homology informationsnoRNA release from pre-rRNA / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / PeBoW complex / ATP-dependent activity, acting on RNA / rRNA primary transcript binding / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of 5.8S rRNA / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...snoRNA release from pre-rRNA / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / PeBoW complex / ATP-dependent activity, acting on RNA / rRNA primary transcript binding / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of 5.8S rRNA / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / preribosome, small subunit precursor / proteasome binding / Major pathway of rRNA processing in the nucleolus and cytosol / ribosomal large subunit binding / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / preribosome, large subunit precursor / 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 / L13a-mediated translational silencing of Ceruloplasmin expression / ribosomal large subunit export from nucleus / 90S preribosome / ribonucleoprotein complex binding / maturation of LSU-rRNA / nuclear periphery / proteasome complex / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / macroautophagy / small-subunit processome / protein catabolic process / rRNA processing / nuclear envelope / ribosomal small subunit biogenesis / 5S rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / protein-macromolecule adaptor activity / cytosolic large ribosomal subunit / cytoplasmic translation / RNA helicase activity / negative regulation of translation / rRNA binding / structural constituent of ribosome / RNA helicase / ribosome / mRNA binding / nucleolus / ATP hydrolysis activity / DNA binding / RNA binding / zinc ion binding / nucleoplasm / ATP binding / identical protein binding / nucleus / cytoplasm / cytosol Similarity search - Function | ||||||
| Biological species | ![]() | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||
Authors | Kater, L. / Cheng, J. / Barrio-Garcia, C. / Hurt, E. / Beckmann, R. | ||||||
Citation | Journal: Cell / Year: 2017Title: Visualizing the Assembly Pathway of Nucleolar Pre-60S Ribosomes. Authors: Lukas Kater / Matthias Thoms / Clara Barrio-Garcia / Jingdong Cheng / Sherif Ismail / Yasar Luqman Ahmed / Gert Bange / Dieter Kressler / Otto Berninghausen / Irmgard Sinning / Ed Hurt / Roland Beckmann / ![]() Abstract: Eukaryotic 60S ribosomal subunits are comprised of three rRNAs and ∼50 ribosomal proteins. The initial steps of their formation take place in the nucleolus, but, owing to a lack of structural ...Eukaryotic 60S ribosomal subunits are comprised of three rRNAs and ∼50 ribosomal proteins. The initial steps of their formation take place in the nucleolus, but, owing to a lack of structural information, this process is poorly understood. Using cryo-EM, we solved structures of early 60S biogenesis intermediates at 3.3 Å to 4.5 Å resolution, thereby providing insights into their sequential folding and assembly pathway. Besides revealing distinct immature rRNA conformations, we map 25 assembly factors in six different assembly states. Notably, the Nsa1-Rrp1-Rpf1-Mak16 module stabilizes the solvent side of the 60S subunit, and the Erb1-Ytm1-Nop7 complex organizes and connects through Erb1's meandering N-terminal extension, eight assembly factors, three ribosomal proteins, and three 25S rRNA domains. Our structural snapshots reveal the order of integration and compaction of the six major 60S domains within early nucleolar 60S particles developing stepwise from the solvent side around the exit tunnel to the central protuberance. | ||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 6em3.cif.gz | 1.3 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb6em3.ent.gz | 833 KB | Display | PDB format |
| PDBx/mmJSON format | 6em3.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 6em3_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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| Full document | 6em3_full_validation.pdf.gz | 1.3 MB | Display | |
| Data in XML | 6em3_validation.xml.gz | 127 KB | Display | |
| Data in CIF | 6em3_validation.cif.gz | 234 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/em/6em3 ftp://data.pdbj.org/pub/pdb/validation_reports/em/6em3 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 3888MC ![]() 3889C ![]() 3890C ![]() 3891C ![]() 3892C ![]() 3893C ![]() 6elzC ![]() 6em1C ![]() 6em4C ![]() 6em5C ![]() 6emfC ![]() 6emgC ![]() 6en7C M: map data used to model this data C: citing same article ( |
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| Similar structure data |
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Assembly
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Components
-Protein , 8 types, 8 molecules x34JvDKn
| #1: Protein | Mass: 35184.387 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P38805 |
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| #3: Protein | Mass: 35754.605 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P10962 |
| #4: Protein | Mass: 33250.531 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P35178 |
| #7: Protein | Mass: 49842.613 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P36049 |
| #8: Protein | Mass: 26954.447 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P40007 |
| #27: Protein | Mass: 56798.348 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: Q03532, RNA helicase |
| #28: Protein | Mass: 42596.691 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P38779 |
| #29: Protein | Mass: 69984.148 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P53261 |
-60S ribosomal protein ... , 17 types, 17 molecules FCeEfGhijLMNOPQSY
| #2: Protein | Mass: 27686.281 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P05737 |
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| #9: Protein | Mass: 39159.125 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P10664 |
| #10: Protein | Mass: 14809.441 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P38061 |
| #11: Protein | Mass: 20000.564 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: Q02326 |
| #12: Protein | Mass: 12177.130 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P05744 |
| #13: Protein | Mass: 28175.820 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P17076 |
| #14: Protein | Mass: 13942.640 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P0CX84 |
| #15: Protein | Mass: 11162.285 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: O14455 |
| #16: Protein | Mass: 9877.395 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P49166 |
| #17: Protein | Mass: 22604.164 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: Q12690 |
| #18: Protein | Mass: 15195.066 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P36105 |
| #19: Protein | Mass: 24482.357 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P05748 |
| #20: Protein | Mass: 22247.227 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P26784 |
| #21: Protein | Mass: 20589.518 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P05740 |
| #22: Protein | Mass: 20609.252 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P0CX49 |
| #23: Protein | Mass: 20478.852 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P0CX23 |
| #24: Protein | Mass: 14265.784 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P05743 |
-Ribosome biogenesis protein ... , 5 types, 5 molecules 5AotB
| #5: Protein | Mass: 51982.457 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P53136 |
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| #6: Protein | Mass: 33585.414 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: Q08235 |
| #30: Protein | Mass: 25499.186 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P53927 |
| #31: Protein | Mass: 36621.074 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: P40693 |
| #33: Protein | Mass: 91830.609 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: UniProt: Q04660 |
-RNA chain , 1 types, 1 molecules 1
| #25: RNA chain | Mass: 1097493.875 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: GenBank: 834774822 |
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-5.8S ribosomal ... , 2 types, 2 molecules 26
| #26: RNA chain | Mass: 50682.922 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: GenBank: 1214879358 |
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| #32: RNA chain | Mass: 74308.391 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) ![]() References: GenBank: 1102641490 |
-Details
| Has protein modification | 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 |
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Sample preparation
| Component | Name: Nsa1-TAP Flag-Ytm1 / Type: RIBOSOME Details: Pre-60S assembly intermediates purified via Nsa1-TAP Flag-Ytm1 Entity ID: all / Source: NATURAL |
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| Source (natural) | Organism: ![]() |
| Buffer solution | pH: 7.5 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| 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 / Cs: 2.7 mm |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Electron dose: 27 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON II (4k x 4k) |
| Image scans | Sampling size: 14 µm / Width: 4096 / Height: 4096 / Movie frames/image: 10 / Used frames/image: 1-6 |
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Processing
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| CTF correction | Type: NONE | ||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 34453 / Symmetry type: POINT | ||||||||||||||||||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT / Space: REAL / Details: Backbone model |
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