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Yorodumi- PDB-5apn: Structure of the yeast 60S ribosomal subunit in complex with Arx1... -
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-Basic information
Entry | Database: PDB / ID: 5apn | ||||||
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Title | Structure of the yeast 60S ribosomal subunit in complex with Arx1, Alb1 and N-terminally tagged Rei1 | ||||||
Components |
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Keywords | RIBOSOME / EUKARYOTIC RIBOSOME BIOGENESIS / RIBOSOME MATURATION / RIBOSOME BIOGENESIS FACTOR / 60S RIBOSOMAL SUBUNIT / REI1 / ARX1 / ALB1 / CRYO-EM / RIBOSOMAL POLYPEPTIDE EXIT TUNNEL PROOFREADING | ||||||
Function / homology | Function and homology information budding cell bud growth / Hydrolases / nucleocytoplasmic transport / 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) / response to cycloheximide / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) ...budding cell bud growth / Hydrolases / nucleocytoplasmic transport / 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) / response to cycloheximide / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / 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 / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / 90S preribosome / regulation of translational fidelity / protein-RNA complex assembly / translational termination / maturation of LSU-rRNA / Neutrophil degranulation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosomal large subunit biogenesis / translational initiation / macroautophagy / maintenance of translational fidelity / modification-dependent protein catabolic process / protein tag activity / rRNA processing / metallopeptidase activity / ribosome biogenesis / viral capsid / mitotic cell cycle / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / sequence-specific DNA binding / negative regulation of translation / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / translation / response to antibiotic / mRNA binding / ubiquitin protein ligase binding / host cell nucleus / nucleolus / proteolysis / RNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||
Biological species | Saccharomyces cerevisiae S288c (yeast) Saccharomyces cerevisiae S288C (yeast) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.91 Å | ||||||
Authors | Greber, B.J. / Gerhardy, S. / Leitner, A. / Leibundgut, M. / Salem, M. / Boehringer, D. / Leulliot, N. / Aebersold, R. / Panse, V.G. / Ban, V. | ||||||
Citation | Journal: Cell / Year: 2016 Title: Insertion of the Biogenesis Factor Rei1 Probes the Ribosomal Tunnel during 60S Maturation. Authors: Basil Johannes Greber / Stefan Gerhardy / Alexander Leitner / Marc Leibundgut / Michèle Salem / Daniel Boehringer / Nicolas Leulliot / Ruedi Aebersold / Vikram Govind Panse / Nenad Ban / Abstract: Eukaryotic ribosome biogenesis depends on several hundred assembly factors to produce functional 40S and 60S ribosomal subunits. The final phase of 60S subunit biogenesis is cytoplasmic maturation, ...Eukaryotic ribosome biogenesis depends on several hundred assembly factors to produce functional 40S and 60S ribosomal subunits. The final phase of 60S subunit biogenesis is cytoplasmic maturation, which includes the proofreading of functional centers of the 60S subunit and the release of several ribosome biogenesis factors. We report the cryo-electron microscopy (cryo-EM) structure of the yeast 60S subunit in complex with the biogenesis factors Rei1, Arx1, and Alb1 at 3.4 Å resolution. In addition to the network of interactions formed by Alb1, the structure reveals a mechanism for ensuring the integrity of the ribosomal polypeptide exit tunnel. Arx1 probes the entire set of inner-ring proteins surrounding the tunnel exit, and the C terminus of Rei1 is deeply inserted into the ribosomal tunnel, where it forms specific contacts along almost its entire length. We provide genetic and biochemical evidence that failure to insert the C terminus of Rei1 precludes subsequent steps of 60S maturation. | ||||||
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 | 5apn.cif.gz | 3.2 MB | Display | PDBx/mmCIF format |
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PDB format | pdb5apn.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 5apn.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 5apn_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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Full document | 5apn_full_validation.pdf.gz | 1.3 MB | Display | |
Data in XML | 5apn_validation.xml.gz | 203.5 KB | Display | |
Data in CIF | 5apn_validation.cif.gz | 362.3 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ap/5apn ftp://data.pdbj.org/pub/pdb/validation_reports/ap/5apn | HTTPS FTP |
-Related structure data
Related structure data | 3152MC 3151C 3153C 5apoC C: citing same article (ref.) M: map data used to model this data |
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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 578
#1: RNA chain | Mass: 1094369.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Saccharomyces cerevisiae S288c (yeast) / References: GenBank: 329138943 |
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#2: RNA chain | Mass: 38951.105 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Saccharomyces cerevisiae S288c (yeast) / References: GenBank: 329138943 |
#3: RNA chain | Mass: 50682.922 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Saccharomyces cerevisiae S288c (yeast) / References: GenBank: 329138943 |
+60S ribosomal protein ... , 39 types, 39 molecules ABCDEFGHIJLMNOPQRSTUVWXYZabcde...
-Protein , 5 types, 5 molecules mqxyz
#41: Protein | Mass: 14583.077 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Saccharomyces cerevisiae S288c (yeast) / References: UniProt: P0CH08 |
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#44: Protein | Mass: 33749.121 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Saccharomyces cerevisiae S288c (yeast) / References: UniProt: P05317 |
#45: Protein | Mass: 67835.188 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae S288c (yeast) / Production host: Escherichia coli BL21(DE3) (bacteria) / Variant (production host): ROSETTA 2 / References: UniProt: Q03862, Hydrolases |
#46: Protein | Mass: 48492.180 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae S288c (yeast) / Plasmid: PPROEX-HTB / Production host: Escherichia coli BL21(DE3) (bacteria) / Variant (production host): ROSETTA 2 / References: UniProt: P38344 |
#47: Protein | Mass: 7251.930 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae S288C (yeast) / Production host: Escherichia coli BL21(DE3) (bacteria) / Variant (production host): ROSETTA 2 |
-Non-polymers , 2 types, 286 molecules
#48: Chemical | ChemComp-MG / #49: Chemical | ChemComp-ZN / |
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-Details
Has protein modification | N |
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Sequence details | The correct register of the chain z remains unknown for some parts of the model. Hence they are ...The correct register of the chain z remains unknown for some parts of the model. Hence they are built in as UNK. However, the sample sequence for chain z corresponds to UniProt P47019.1 |
-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: YEAST 60S ARX1 ALB1 6XHIS-REI1 / Type: RIBOSOME Details: QUANTIFOIL HOLEY CARBON GRIDS WERE COATED WITH A THIN CARBON FILM |
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Buffer solution | Name: 20MM / pH: 8 / Details: 20MM |
Specimen | Conc.: 0.2 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Details: HOLEY CARBON |
Vitrification | Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE-PROPANE / Details: PLUNGE-FROZEN |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS / Date: Apr 17, 2015 / Details: COLLECTED IN MOVIE MODE |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 59000 X / Calibrated magnification: 100720 X / Nominal defocus max: 3000 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm |
Specimen holder | Temperature: 80 K |
Image recording | Electron dose: 20 e/Å2 / Film or detector model: FEI FALCON II (4k x 4k) |
Image scans | Num. digital images: 670 |
-Processing
EM software |
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CTF correction | Details: PER DETECTOR FRAME | ||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||
3D reconstruction | Method: MAXIMUM LIKELIHOOD BASED REFINEMENT IMPLEMENTED IN RELION Resolution: 3.91 Å / Num. of particles: 22040 / Actual pixel size: 1.39 Å Details: FOR VISUALIZATION PURPOSES THE FINAL MAP WAS FILTERED AND AMPLITUDE CORRECTED IN RELION. SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-3152 (DEPOSITION ID 13746). THE COORDINATES WERE ...Details: FOR VISUALIZATION PURPOSES THE FINAL MAP WAS FILTERED AND AMPLITUDE CORRECTED IN RELION. SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-3152 (DEPOSITION ID 13746). THE COORDINATES WERE REFINED IN RECIPROCAL SPACE USING PHENIX.REFINE AGAINST THE MLHL TARGET. FOR THIS, THE CRYO-EM MAP WAS CONVERTED TO RECIPROCAL SPACE STRUCTURE FACTORS. Symmetry type: POINT | ||||||||||||||||||||
Atomic model building | Space: REAL |