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Open data
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Basic information
| Entry | Database: PDB / ID: 9e0n | ||||||
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| Title | M. smegmatis unmethylated 70S ribosome structure | ||||||
Components |
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Keywords | RIBOSOME / 70S ribosome / unmethylated ribosome / ribonucleoprotein complex / protein synthesis | ||||||
| Function / homology | Function and homology informationlarge ribosomal subunit / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / 5S rRNA binding / ribosomal large subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit ...large ribosomal subunit / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / 5S rRNA binding / ribosomal large subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / mRNA binding / RNA binding / zinc ion binding / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||
| Biological species | Mycolicibacterium smegmatis (bacteria) | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.24 Å | ||||||
Authors | Nandi, S. / Conn, G.L. | ||||||
| Funding support | United States, 1items
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Citation | Journal: Nucleic Acids Res / Year: 2025Title: Distant ribose 2'-O-methylation of 23S rRNA helix 69 pre-orders the capreomycin drug binding pocket at the ribosome subunit interface. Authors: Suparno Nandi / Debayan Dey / Pooja Srinivas / Christine M Dunham / Graeme L Conn / ![]() Abstract: Loss of ribosomal RNA (rRNA) modifications incorporated by the intrinsic methyltransferase TlyA results in reduced sensitivity to tuberactinomycin antibiotics such as capreomycin. However, how rRNA ...Loss of ribosomal RNA (rRNA) modifications incorporated by the intrinsic methyltransferase TlyA results in reduced sensitivity to tuberactinomycin antibiotics such as capreomycin. However, how rRNA methylation alters drug binding, particularly at the distant but functionally more important site in 23S rRNA helix 69 (H69), is currently unknown. We determined high-resolution cryo-electron microscopy structures of the Mycolicibacterium smegmatis 70S ribosome with or without the two ribose 2'-O-methyl modifications incorporated by TlyA. In the unmodified ribosome, the tip of H69 adopts a more compact conformation, positioning two key nucleotides (A2137 and C2138) such that interactions with capreomycin would be lost and the binding pocket partially occluded. Methylation of 23S rRNA nucleotide C2144 promotes conformational changes that result in a more favorable positioning of C2138 and adoption of a more open conformation to enable capreomycin binding. Molecular dynamics simulations and H69 RNA helical analyses additionally reveal specific propagation of these changes from the site of modification to the H69 tip, allosterically reconfiguring the capreomycin binding site. Methylation of h44 also results in structural rearrangements at the H69-h44 interface to support maintenance of these changes that favor antibiotic binding. This work thus reveals the effect and regulation of distant rRNA methylation on ribosome-targeting antibiotic binding. | ||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 9e0n.cif.gz | 3.2 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb9e0n.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 9e0n.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 9e0n_validation.pdf.gz | 1.9 MB | Display | wwPDB validaton report |
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| Full document | 9e0n_full_validation.pdf.gz | 2 MB | Display | |
| Data in XML | 9e0n_validation.xml.gz | 239.1 KB | Display | |
| Data in CIF | 9e0n_validation.cif.gz | 402.1 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/e0/9e0n ftp://data.pdbj.org/pub/pdb/validation_reports/e0/9e0n | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 47363MC ![]() 9e0pC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
+Large ribosomal subunit protein ... , 29 types, 29 molecules 123456CDEFGIJKLMNOPQRSTUVWXYZ
-50S ribosomal protein ... , 3 types, 3 molecules 78H
| #7: Protein/peptide | Mass: 4341.277 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSL4 |
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| #8: Protein/peptide | Mass: 2841.375 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QTP4 |
| #16: Protein | Mass: 15949.253 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0A653F9J9 |
-RNA chain , 3 types, 3 molecules ABa
| #9: RNA chain | Mass: 1012140.938 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: GenBank: 2198294598 |
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| #10: RNA chain | Mass: 38038.777 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: GenBank: 2093960070 |
| #35: RNA chain | Mass: 495373.656 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: GenBank: 2093960070 |
-Small ribosomal subunit protein ... , 19 types, 19 molecules bcdefghijklmnopqrst
| #36: Protein | Mass: 30145.230 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QVB8 |
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| #37: Protein | Mass: 30191.227 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSD7 |
| #38: Protein | Mass: 23415.787 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSL7 |
| #39: Protein | Mass: 21946.090 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSG6 |
| #40: Protein | Mass: 10991.637 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0A2U9Q0X2 |
| #41: Protein | Mass: 17660.375 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QS97 |
| #42: Protein | Mass: 14492.638 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSG3 |
| #43: Protein | Mass: 16794.365 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSP9 |
| #44: Protein | Mass: 11454.313 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSD0 |
| #45: Protein | Mass: 14671.762 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSL6 |
| #46: Protein | Mass: 13896.366 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QS96 |
| #47: Protein | Mass: 14249.619 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSL5 |
| #48: Protein | Mass: 6976.409 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSG2 |
| #49: Protein | Mass: 10368.097 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QVQ3 |
| #50: Protein | Mass: 16795.207 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QV37 |
| #51: Protein | Mass: 11127.002 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSE0 |
| #52: Protein | Mass: 9524.188 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0R7F7 |
| #53: Protein | Mass: 10800.602 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QSD5 |
| #54: Protein | Mass: 9556.104 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0R102 |
-Protein/peptide / Non-polymers , 2 types, 108 molecules u

| #55: Protein/peptide | Mass: 4164.300 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A / References: UniProt: A0QR10 |
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| #56: Chemical | ChemComp-MG / |
-Details
| Has ligand of interest | N |
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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 |
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Sample preparation
| Component | Name: M. smegmatis unmethylated 70S ribosome / Type: RIBOSOME / Entity ID: #1-#55 / Source: NATURAL |
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| Molecular weight | Value: 2.3 MDa / Experimental value: NO |
| Source (natural) | Organism: Mycolicibacterium smegmatis (bacteria) / Strain: LR222 C101A |
| Buffer solution | pH: 7.6 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: TFS KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2100 nm / Nominal defocus min: 600 nm / Cs: 2.7 mm |
| Image recording | Average exposure time: 2 sec. / Electron dose: 52.51 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of real images: 5060 |
| Image scans | Width: 5760 / Height: 4092 |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.24 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 227969 / Symmetry type: POINT | ||||||||||||||||||||||||
| Atomic model building | Protocol: FLEXIBLE FIT | ||||||||||||||||||||||||
| Atomic model building | PDB-ID: 5ZEB Accession code: 5ZEB / Source name: PDB / Type: experimental model |
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Mycolicibacterium smegmatis (bacteria)
United States, 1items
Citation


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FIELD EMISSION GUN
