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Yorodumi- PDB-7qh7: Cryo-EM structure of the human mtLSU assembly intermediate upon M... -
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Basic information
| Entry | Database: PDB / ID: 7qh7 | ||||||
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| Title | Cryo-EM structure of the human mtLSU assembly intermediate upon MRM2 depletion - class 4 | ||||||
|  Components | 
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|  Keywords | RIBOSOME / Mitochondria / Assembly / Methyltransferase / MRM2 / RNA modification | ||||||
| Function / homology |  Function and homology information negative regulation of mitochondrial translation / mitochondrial large ribosomal subunit assembly / protein lipoylation / negative regulation of ribosome biogenesis / Complex I biogenesis / Mitochondrial Fatty Acid Beta-Oxidation / Protein lipoylation / positive regulation of mitochondrial translation / Respiratory electron transport / rRNA import into mitochondrion ...negative regulation of mitochondrial translation / mitochondrial large ribosomal subunit assembly / protein lipoylation / negative regulation of ribosome biogenesis / Complex I biogenesis / Mitochondrial Fatty Acid Beta-Oxidation / Protein lipoylation / positive regulation of mitochondrial translation / Respiratory electron transport / rRNA import into mitochondrion / mitochondrial translational elongation / Mitochondrial translation elongation / Mitochondrial translation termination / Mitochondrial translation initiation / mitochondrial [2Fe-2S] assembly complex / iron-sulfur cluster assembly complex / mitochondrial large ribosomal subunit / mitochondrial fission / mitochondrial large ribosomal subunit binding / mitochondrial ribosome / mitochondrial small ribosomal subunit / Hydrolases; Acting on ester bonds; Endoribonucleases producing 5'-phosphomonoesters / mitochondrial translation / [2Fe-2S] cluster assembly / iron-sulfur cluster assembly / acyl binding / acyl carrier activity / ribosomal large subunit binding / mitochondrial respiratory chain complex I assembly / mitochondrial electron transport, NADH to ubiquinone / proton motive force-driven mitochondrial ATP synthesis / respiratory chain complex I / RNA processing / Mitochondrial protein degradation / aerobic respiration / cellular response to leukemia inhibitory factor / fatty acid binding / ribosomal large subunit biogenesis / mitochondrial membrane / fatty acid biosynthetic process / double-stranded RNA binding / 5S rRNA binding / small ribosomal subunit rRNA binding / endonuclease activity / mitochondrial inner membrane / negative regulation of translation / rRNA binding / nuclear body / structural constituent of ribosome / ribosome / translation / mitochondrial matrix / ribonucleoprotein complex / protein domain specific binding / nucleotide binding / mRNA binding / apoptotic process / calcium ion binding / structural molecule activity / mitochondrion / extracellular space / RNA binding / nucleoplasm / nucleus / plasma membrane / cytosol Similarity search - Function | ||||||
| Biological species |  Homo sapiens (human) | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.89 Å | ||||||
|  Authors | Rebelo-Guiomar, P. / Pellegrino, S. / Dent, K.C. / Warren, A.J. / Minczuk, M. | ||||||
| Funding support |  United Kingdom, 1items 
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|  Citation |  Journal: Nat Commun / Year: 2022 Title: A late-stage assembly checkpoint of the human mitochondrial ribosome large subunit. Authors: Pedro Rebelo-Guiomar / Simone Pellegrino / Kyle C Dent / Aldema Sas-Chen / Leonor Miller-Fleming / Caterina Garone / Lindsey Van Haute / Jack F Rogan / Adam Dinan / Andrew E Firth / Byron ...Authors: Pedro Rebelo-Guiomar / Simone Pellegrino / Kyle C Dent / Aldema Sas-Chen / Leonor Miller-Fleming / Caterina Garone / Lindsey Van Haute / Jack F Rogan / Adam Dinan / Andrew E Firth / Byron Andrews / Alexander J Whitworth / Schraga Schwartz / Alan J Warren / Michal Minczuk /      Abstract: Many cellular processes, including ribosome biogenesis, are regulated through post-transcriptional RNA modifications. Here, a genome-wide analysis of the human mitochondrial transcriptome shows that ...Many cellular processes, including ribosome biogenesis, are regulated through post-transcriptional RNA modifications. Here, a genome-wide analysis of the human mitochondrial transcriptome shows that 2'-O-methylation is limited to residues of the mitoribosomal large subunit (mtLSU) 16S mt-rRNA, introduced by MRM1, MRM2 and MRM3, with the modifications installed by the latter two proteins being interdependent. MRM2 controls mitochondrial respiration by regulating mitoribosome biogenesis. In its absence, mtLSU particles (visualized by cryo-EM at the resolution of 2.6 Å) present disordered RNA domains, partial occupancy of bL36m and bound MALSU1:L0R8F8:mtACP anti-association module, allowing five mtLSU biogenesis intermediates with different intersubunit interface configurations to be placed along the assembly pathway. However, mitoribosome biogenesis does not depend on the methyltransferase activity of MRM2. Disruption of the MRM2 Drosophila melanogaster orthologue leads to mitochondria-related developmental arrest. This work identifies a key checkpoint during mtLSU assembly, essential to maintain mitochondrial homeostasis. | ||||||
| History | 
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- Structure visualization
Structure visualization
| Structure viewer | Molecule:  Molmil  Jmol/JSmol | 
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- Downloads & links
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- Download
Download
| PDBx/mmCIF format |  7qh7.cif.gz | 1.8 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb7qh7.ent.gz | 1.5 MB | Display |  PDB format | 
| PDBx/mmJSON format |  7qh7.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  7qh7_validation.pdf.gz | 1.1 MB | Display |  wwPDB validaton report | 
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| Full document |  7qh7_full_validation.pdf.gz | 1.1 MB | Display | |
| Data in XML |  7qh7_validation.xml.gz | 174.2 KB | Display | |
| Data in CIF |  7qh7_validation.cif.gz | 301.6 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/qh/7qh7  ftp://data.pdbj.org/pub/pdb/validation_reports/qh/7qh7 | HTTPS FTP | 
-Related structure data
| Related structure data |  13967MC  7qh6C C: citing same article ( M: map data used to model this data | 
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| Similar structure data | Similarity search - Function & homology  F&H Search | 
- Links
Links
- Assembly
Assembly
| Deposited unit |  
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| 1 | 
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- Components
Components
+39S ribosomal protein  ... , 41 types, 41 molecules DEFHIKLMNOPQRSTUVWXYZ01235679a...                              
-Protein , 6 types, 6 molecules opquvw     
| #39: Protein | Mass: 9768.266 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / References: UniProt: Q9BQC6 | 
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| #40: Protein | Mass: 14901.154 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) | 
| #41: Protein | Mass: 11942.498 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / References: UniProt: Q8TAE8 | 
| #44: Protein | Mass: 13280.472 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / References: UniProt: Q96EH3 | 
| #45: Protein | Mass: 8329.592 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / References: UniProt: L0R8F8 | 
| #46: Protein | Mass: 9144.511 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) / References: UniProt: O14561 | 
-RNA chain , 2 types, 2 molecules AB 
| #47: RNA chain | Mass: 403061.188 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) | 
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| #48: RNA chain | Mass: 19530.658 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)   Homo sapiens (human) | 
-Non-polymers , 3 types, 67 molecules 




| #50: Chemical | ChemComp-MG / #51: Chemical | #52: Water | ChemComp-HOH / |  | 
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-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 | 
- Sample preparation
Sample preparation
| Component | Name: Human mitochondrial ribosome large subunit / Type: RIBOSOME / Entity ID: #1-#49 / Source: NATURAL | 
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| Molecular weight | Experimental value: NO | 
| Source (natural) | Organism:  Homo sapiens (human) | 
| Buffer solution | pH: 7.4 | 
| 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 R2/2 | 
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K | 
- Electron microscopy imaging
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: 2600 nm / Nominal defocus min: 1000 nm | 
| Image recording | Electron dose: 52.5 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON III (4k x 4k) | 
- Processing
Processing
| EM software | 
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 2709521 | ||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 2.89 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 224933 / Num. of class averages: 1 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT / Space: REAL | ||||||||||||||||||||||||||||||||||||
| Atomic model building | PDB-ID: 5OOL Accession code: 5OOL / Source name: PDB / Type: experimental model | 
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