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Yorodumi- PDB-8a98: CRYO-EM STRUCTURE OF LEISHMANIA MAJOR 80S RIBOSOME : snoRNA MUTANT -
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-Basic information
Entry | Database: PDB / ID: 8a98 | ||||||
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Title | CRYO-EM STRUCTURE OF LEISHMANIA MAJOR 80S RIBOSOME : snoRNA MUTANT | ||||||
Components |
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Keywords | RIBOSOME / CRYO-EM / LEISHMANIA MAJOR / 80S RIBOSOME snoRNA MUTANT | ||||||
Function / homology | Function and homology information ciliary transition zone / nuclear lumen / ciliary plasm / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / protein-RNA complex assembly / translation regulator activity / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rescue of stalled ribosome ...ciliary transition zone / nuclear lumen / ciliary plasm / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / protein-RNA complex assembly / translation regulator activity / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rescue of stalled ribosome / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / protein kinase C binding / maintenance of translational fidelity / rRNA processing / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosome biogenesis / ribosome binding / kinase activity / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / positive regulation of protein phosphorylation / ribonucleoprotein complex / translation / mRNA binding / nucleolus / RNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||
Biological species | Leishmania major strain Friedlin (eukaryote) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.46 Å | ||||||
Authors | Rajan, K.S. / Yonath, A. / Bashan, A. | ||||||
Funding support | Israel, 1items
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Citation | Journal: Cell Rep / Year: 2024 Title: Structural and mechanistic insights into the function of Leishmania ribosome lacking a single pseudouridine modification. Authors: K Shanmugha Rajan / Saurav Aryal / Disha-Gajanan Hiregange / Anat Bashan / Hava Madmoni / Mika Olami / Tirza Doniger / Smadar Cohen-Chalamish / Pascal Pescher / Masato Taoka / Yuko Nobe / ...Authors: K Shanmugha Rajan / Saurav Aryal / Disha-Gajanan Hiregange / Anat Bashan / Hava Madmoni / Mika Olami / Tirza Doniger / Smadar Cohen-Chalamish / Pascal Pescher / Masato Taoka / Yuko Nobe / Aliza Fedorenko / Tanaya Bose / Ella Zimermann / Eric Prina / Noa Aharon-Hefetz / Yitzhak Pilpel / Toshiaki Isobe / Ron Unger / Gerald F Späth / Ada Yonath / Shulamit Michaeli / Abstract: Leishmania is the causative agent of cutaneous and visceral diseases affecting millions of individuals worldwide. Pseudouridine (Ψ), the most abundant modification on rRNA, changes during the ...Leishmania is the causative agent of cutaneous and visceral diseases affecting millions of individuals worldwide. Pseudouridine (Ψ), the most abundant modification on rRNA, changes during the parasite life cycle. Alterations in the level of a specific Ψ in helix 69 (H69) affected ribosome function. To decipher the molecular mechanism of this phenotype, we determine the structure of ribosomes lacking the single Ψ and its parental strain at ∼2.4-3 Å resolution using cryo-EM. Our findings demonstrate the significance of a single Ψ on H69 to its structure and the importance for its interactions with helix 44 and specific tRNAs. Our study suggests that rRNA modification affects translation of mRNAs carrying codon bias due to selective accommodation of tRNAs by the ribosome. Based on the high-resolution structures, we propose a mechanism explaining how the ribosome selects specific tRNAs. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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PDBx/mmCIF format | 8a98.cif.gz | 4.9 MB | Display | PDBx/mmCIF format |
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PDB format | pdb8a98.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 8a98.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8a98_validation.pdf.gz | 1.9 MB | Display | wwPDB validaton report |
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Full document | 8a98_full_validation.pdf.gz | 2.2 MB | Display | |
Data in XML | 8a98_validation.xml.gz | 329 KB | Display | |
Data in CIF | 8a98_validation.cif.gz | 587.8 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/a9/8a98 ftp://data.pdbj.org/pub/pdb/validation_reports/a9/8a98 | HTTPS FTP |
-Related structure data
Related structure data | 15272MC 8ovjC 8rxhC 8rxxC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-Putative 40S ribosomal protein ... , 15 types, 15 molecules SWSYSVSUSTSRSCSDScSPSNSdSLSJK
#1: Protein | Mass: 17472.395 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QBV0 |
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#3: Protein | Mass: 17035.592 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QH01 |
#5: Protein | Mass: 16551.529 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q806 |
#6: Protein | Mass: 20023.457 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
#7: Protein | Mass: 17482.764 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q3M1 |
#9: Protein | Mass: 17433.422 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q1Y3 |
#11: Protein | Mass: 24511.762 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: BPK282A1 / References: UniProt: Q4Q4A0 |
#12: Protein | Mass: 22188.902 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QIM3 |
#18: Protein | Mass: 9819.463 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: BPK282A1 / References: UniProt: Q4Q140 |
#22: Protein | Mass: 15969.643 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: BPK282A1 / References: UniProt: Q4QC89 |
#24: Protein | Mass: 18681.125 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q1X7 |
#25: Protein | Mass: 9763.191 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q931 |
#27: Protein | Mass: 16718.586 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: BPK282A1 / References: UniProt: Q4Q9A5 |
#28: Protein | Mass: 14719.258 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QGW3 |
#61: Protein | Mass: 19948.615 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QBJ1 |
-40S ribosomal protein ... , 15 types, 15 molecules SZSXSBSESKSaSeSQSOSISHSGSFSASb
#2: Protein | Mass: 15808.882 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q1D2 |
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#4: Protein | Mass: 18144.053 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: E9AEE8 |
#10: Protein | Mass: 27536.629 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q0Q0 |
#13: Protein | Mass: 30742.084 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QG31 |
#14: Protein | Mass: 24984.250 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: P25204 |
#17: Protein | Mass: 13071.399 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q9X4 |
#19: Protein | Mass: 7394.894 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q7P0 |
#21: Protein | Mass: 15627.837 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QG97 |
#23: Protein | Mass: 15623.180 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q8H1 |
#29: Protein | Mass: 23765.207 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: E9AC32 |
#30: Protein | Mass: 21336.623 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q868B1 |
#31: Protein | Mass: 28370.227 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q9NE83 |
#32: Protein | Mass: 28791.135 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QDL5 |
#33: Protein | Mass: 30062.037 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4FX73 |
#34: Protein | Mass: 12794.251 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q8L6 |
-Putative ribosomal protein ... , 7 types, 7 molecules SSSMBgXkC
#8: Protein | Mass: 6696.929 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q817 |
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#26: Protein | Mass: 13031.983 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: BPK282A1 / References: UniProt: Q4Q8G4 |
#45: Protein | Mass: 47593.660 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q2Q6 |
#68: Protein | Mass: 16492.312 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QHL3 |
#76: Protein | Mass: 14608.168 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q1W6 |
#77: Protein | Mass: 9479.287 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: BPK282A1 / References: UniProt: Q4Q8X0 |
#80: Protein | Mass: 41096.855 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: E9ADW5 |
-RNA chain , 10 types, 10 molecules S4S112478563
#15: RNA chain | Mass: 6106.721 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
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#20: RNA chain | Mass: 710459.375 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
#41: RNA chain | Mass: 577004.438 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
#42: RNA chain | Mass: 492524.812 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
#43: RNA chain | Mass: 59144.129 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: GenBank: 321438308 |
#44: RNA chain | Mass: 54952.504 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: GenBank: 1207899567 |
#46: RNA chain | Mass: 39451.383 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
#47: RNA chain | Mass: 43481.770 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: GenBank: 321438308 |
#58: RNA chain | Mass: 23336.938 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: GenBank: 321438308 |
#81: RNA chain | Mass: 69146.633 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: GenBank: 321438308 |
-Protein , 3 types, 3 molecules SgShI
#16: Protein | Mass: 34441.516 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q7Y7 |
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#35: Protein | Mass: 25229.275 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q5K7 |
#53: Protein | Mass: 24805.891 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: E9AEA8 |
-60S ribosomal protein ... , 10 types, 10 molecules fdbGPRDYon
#36: Protein | Mass: 15458.153 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QC11 |
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#37: Protein | Mass: 11370.224 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: E9AEK1 |
#40: Protein | Mass: 8001.308 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) Strain: MHOM/GT/2001/U1103 / References: UniProt: Q4Q178 |
#50: Protein | Mass: 29824.182 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QIP1 |
#54: Protein | Mass: 22071.080 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QG98 |
#56: Protein | Mass: 20852.338 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q5J4 |
#62: Protein | Mass: 21694.502 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: P48157 |
#64: Protein | Mass: 15469.330 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q504 |
#75: Protein | Mass: 10337.223 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4QC01 |
#79: Protein/peptide | Mass: 4390.496 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) |
+Putative 60S ... , 21 types, 21 molecules caAOHQETSLZFhJVWUiple
-Ribosomal protein ... , 2 types, 2 molecules Mj
#52: Protein | Mass: 24497.971 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: Q4Q6S3 |
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#83: Protein | Mass: 9862.590 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Leishmania major strain Friedlin (eukaryote) References: UniProt: E9AEH2 |
-Non-polymers , 5 types, 2431 molecules
#84: Chemical | ChemComp-MG / #85: Chemical | ChemComp-ZN / #86: Chemical | ChemComp-NA / #87: Chemical | #88: Water | ChemComp-HOH / | |
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-Details
Has ligand of interest | Y |
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Has protein modification | Y |
-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: CRYO-EM STRUCTURE OF LEISHMANIA MAJOR 80S RIBOSOME snoRNA MUTANT Type: RIBOSOME Details: CRYO-EM STRUCTURE OF LEISHMANIA MAJOR 80S RIBOSOME snoRNA MUTANT Entity ID: #1-#83 / Source: NATURAL | ||||||||||||||||||||||||
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Source (natural) | Organism: Leishmania major strain Friedlin (eukaryote) | ||||||||||||||||||||||||
Buffer solution | pH: 7.6 | ||||||||||||||||||||||||
Buffer component |
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Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||
Specimen support | Grid material: COPPER / Grid type: Quantifoil R2/2 | ||||||||||||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K |
-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 / Nominal defocus max: 1500 nm / Nominal defocus min: 500 nm |
Image recording | Electron dose: 1 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) |
-Processing
Software | Name: PHENIX / Version: (1.20.1_4487:phenix.real_space_refine) / Classification: refinement | ||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||
3D reconstruction | Resolution: 2.46 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 345376 / Symmetry type: POINT | ||||||||||||||||||||||||||||
Atomic model building |
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Refinement | Cross valid method: THROUGHOUT | ||||||||||||||||||||||||||||
Displacement parameters | Biso max: 271.93 Å2 / Biso mean: 50.2839 Å2 / Biso min: 0 Å2 |