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Yorodumi- PDB-6em5: State D architectural model (Nsa1-TAP Flag-Ytm1) - Visualizing th... -
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-Basic information
Entry | Database: PDB / ID: 6em5 | ||||||
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Title | State D 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 information 25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc2p-Noc3p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / snoRNA release from pre-rRNA / protein-RNA complex remodeling / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity ...25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc2p-Noc3p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / snoRNA release from pre-rRNA / protein-RNA complex remodeling / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (cytosine-C5-)-methyltransferase activity / PeBoW complex / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear pre-replicative complex / rRNA base methylation / rRNA primary transcript binding / ATP-dependent activity, acting on RNA / positive regulation of ATP-dependent activity / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA methylation / maturation of 5.8S rRNA / hexon binding / pre-mRNA 5'-splice site binding / preribosome, small subunit precursor / 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 / 90S preribosome / DNA replication initiation / chromosome organization / 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 / 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 / ribosomal small subunit biogenesis / protein transport / ribosome biogenesis / viral capsid / nuclear envelope / protein-macromolecule adaptor activity / ATPase binding / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / RNA helicase activity / negative regulation of translation / rRNA binding / RNA helicase / ribosome / structural constituent of ribosome / translation / cell division / GTPase activity / mRNA binding / chromatin binding / host cell nucleus / GTP binding / nucleolus / ATP hydrolysis activity / DNA binding / RNA binding / nucleoplasm / ATP binding / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.3 Å | ||||||
Authors | Kater, L. / Cheng, J. / Barrio-Garcia, C. / Hurt, E. / Beckmann, R. | ||||||
Citation | Journal: Cell / Year: 2017 Title: 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. | ||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 6em5.cif.gz | 2.4 MB | Display | PDBx/mmCIF format |
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PDB format | pdb6em5.ent.gz | 1.5 MB | Display | PDB format |
PDBx/mmJSON format | 6em5.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 6em5_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
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Full document | 6em5_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 6em5_validation.xml.gz | 225.9 KB | Display | |
Data in CIF | 6em5_validation.cif.gz | 419.2 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/em/6em5 ftp://data.pdbj.org/pub/pdb/validation_reports/em/6em5 | HTTPS FTP |
-Related structure data
Related structure data | 3890MC 3888C 3889C 3891C 3892C 3893C 6elzC 6em1C 6em3C 6em4C 6emfC 6emgC 6en7C M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-RNA chain , 3 types, 3 molecules 261
#1: RNA chain | Mass: 50682.922 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: GenBank: 1214879358 |
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#2: RNA chain | Mass: 74308.391 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: GenBank: 1102641490 |
#55: RNA chain | Mass: 1097493.875 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: GenBank: 834774822 |
+60S ribosomal protein ... , 29 types, 29 molecules LMNQRSTUZcdefgijkEGOVXYhBCHFP
-Ribosome biogenesis protein ... , 8 types, 8 molecules Amortpu5
#30: Protein | Mass: 33585.414 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q08235 |
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#32: Protein | Mass: 91830.609 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q04660 |
#37: Protein | Mass: 25499.186 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P53927 |
#39: Protein | Mass: 29786.783 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P40078 |
#41: Protein | Mass: 36621.074 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P40693 |
#44: Protein | Mass: 51426.637 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q12024 |
#46: Protein | Mass: 24027.650 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q07915 |
#54: Protein | Mass: 51982.457 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P53136 |
-Protein , 17 types, 17 molecules KDWlbnsyzqvwIJx34
#31: Protein | Mass: 42596.691 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P38779 |
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#33: Protein | Mass: 56798.348 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q03532, RNA helicase |
#34: Protein | Mass: 27098.012 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P33201 |
#35: Protein | Mass: 20411.699 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q08962 |
#36: Protein | Mass: 74531.227 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q02892 |
#38: Protein | Mass: 69984.148 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P53261 |
#40: Protein | Mass: 57798.652 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P40010 |
#42: Protein | Mass: 26476.605 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q12522 |
#43: Protein | Mass: 12435.429 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P38202 |
#45: Protein | Mass: 69912.164 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P40991, 25S rRNA (cytosine2870-C5)-methyltransferase |
#47: Protein | Mass: 26954.447 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P40007 |
#48: Protein | Mass: 96656.172 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P25582, 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase |
#49: Protein | Mass: 75689.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: Q07896 |
#50: Protein | Mass: 49842.613 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P36049 |
#51: Protein | Mass: 35184.387 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P38805 |
#52: Protein | Mass: 35754.605 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P10962 |
#53: Protein | Mass: 33250.531 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: Backbone model Source: (natural) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) References: UniProt: P35178 |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Nsa1-TAP Flag-Ytm1 / Type: RIBOSOME Details: Pre-60S intermediates purified via Nsa1-TAP Flag-Ytm1 Entity ID: all / Source: NATURAL |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Buffer solution | pH: 7.5 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-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 |
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 |
-Processing
EM software |
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CTF correction | Type: NONE | ||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||
3D reconstruction | Resolution: 4.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 37251 / Algorithm: FOURIER SPACE / Symmetry type: POINT | ||||||||||||||||||||||||||||
Atomic model building | Protocol: RIGID BODY FIT / Space: REAL / Details: Backbone model |