+Open data
-Basic information
Entry | Database: PDB / ID: 7pkt | |||||||||||||||
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Title | Large subunit of the Chlamydomonas reinhardtii mitoribosome | |||||||||||||||
Components |
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Keywords | RIBOSOME / Mitochondria / mitoribosome / alga | |||||||||||||||
Function / homology | Function and homology information chloroplast rRNA processing / regulation of mitochondrial mRNA stability / ribonucleoprotein granule / mitochondrial RNA processing / mitochondrial large ribosomal subunit / mitochondrial ribosome / mitochondrial translation / chloroplast / large ribosomal subunit / 5S rRNA binding ...chloroplast rRNA processing / regulation of mitochondrial mRNA stability / ribonucleoprotein granule / mitochondrial RNA processing / mitochondrial large ribosomal subunit / mitochondrial ribosome / mitochondrial translation / chloroplast / large ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / mitochondrial matrix / ribonucleoprotein complex / translation / mRNA binding / mitochondrion / RNA binding / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Chlamydomonas reinhardtii (plant) | |||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3 Å | |||||||||||||||
Authors | Waltz, F. / Soufari, H. / Hashem, Y. | |||||||||||||||
Funding support | European Union, France, 4items
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Citation | Journal: Nat Commun / Year: 2021 Title: How to build a ribosome from RNA fragments in Chlamydomonas mitochondria. Authors: Florent Waltz / Thalia Salinas-Giegé / Robert Englmeier / Herrade Meichel / Heddy Soufari / Lauriane Kuhn / Stefan Pfeffer / Friedrich Förster / Benjamin D Engel / Philippe Giegé / ...Authors: Florent Waltz / Thalia Salinas-Giegé / Robert Englmeier / Herrade Meichel / Heddy Soufari / Lauriane Kuhn / Stefan Pfeffer / Friedrich Förster / Benjamin D Engel / Philippe Giegé / Laurence Drouard / Yaser Hashem / Abstract: Mitochondria are the powerhouse of eukaryotic cells. They possess their own gene expression machineries where highly divergent and specialized ribosomes, named hereafter mitoribosomes, translate the ...Mitochondria are the powerhouse of eukaryotic cells. They possess their own gene expression machineries where highly divergent and specialized ribosomes, named hereafter mitoribosomes, translate the few essential messenger RNAs still encoded by mitochondrial genomes. Here, we present a biochemical and structural characterization of the mitoribosome in the model green alga Chlamydomonas reinhardtii, as well as a functional study of some of its specific components. Single particle cryo-electron microscopy resolves how the Chlamydomonas mitoribosome is assembled from 13 rRNA fragments encoded by separate non-contiguous gene pieces. Additional proteins, mainly OPR, PPR and mTERF helical repeat proteins, are found in Chlamydomonas mitoribosome, revealing the structure of an OPR protein in complex with its RNA binding partner. Targeted amiRNA silencing indicates that these ribosomal proteins are required for mitoribosome integrity. Finally, we use cryo-electron tomography to show that Chlamydomonas mitoribosomes are attached to the inner mitochondrial membrane via two contact points mediated by Chlamydomonas-specific proteins. Our study expands our understanding of mitoribosome diversity and the various strategies these specialized molecular machines adopt for membrane tethering. | |||||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7pkt.cif.gz | 2.3 MB | Display | PDBx/mmCIF format |
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PDB format | pdb7pkt.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 7pkt.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7pkt_validation.pdf.gz | 2.1 MB | Display | wwPDB validaton report |
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Full document | 7pkt_full_validation.pdf.gz | 2.2 MB | Display | |
Data in XML | 7pkt_validation.xml.gz | 219.3 KB | Display | |
Data in CIF | 7pkt_validation.cif.gz | 369.3 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/pk/7pkt ftp://data.pdbj.org/pub/pdb/validation_reports/pk/7pkt | HTTPS FTP |
-Related structure data
Related structure data | 13480MC 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
+Protein , 34 types, 34 molecules abcdjklpqstuvwxyABeDEFIJKLMNOP...
-50S ribosomal protein ... , 2 types, 2 molecules fo
#5: Protein | Mass: 30622.402 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A0A2K3DRF9 |
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#12: Protein | Mass: 13140.527 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8HNL1 |
-Mitochondrial ribosomal protein ... , 6 types, 6 molecules imnrzG
#6: Protein | Mass: 29714.832 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8J810 |
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#10: Protein | Mass: 21380.727 Da / Num. of mol.: 1 / Source method: isolated from a natural source Details: The protein is a mixture of A0A2K3DXS2 and A8JH49,The protein is a mixture of A0A2K3DXS2 and A8JH49 Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8JH49, UniProt: A0A2K3DXS2 |
#11: Protein | Mass: 33412.934 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8I7A8 |
#15: Protein | Mass: 25593.826 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8J6R9 |
#23: Protein | Mass: 6864.234 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8J9C3 |
#30: Protein | Mass: 27320.754 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) / References: UniProt: A8IWI1 |
-Protein/peptide , 3 types, 3 molecules CXZ
#35: Protein/peptide | Mass: 3081.790 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
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#43: Protein/peptide | Mass: 2656.265 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#45: Protein/peptide | Mass: 4188.154 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
-RNA chain , 9 types, 9 molecules 120345678
#46: RNA chain | Mass: 51976.762 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
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#47: RNA chain | Mass: 27512.189 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#48: RNA chain | Mass: 24033.252 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#49: RNA chain | Mass: 59395.223 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#50: RNA chain | Mass: 38253.898 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#51: RNA chain | Mass: 47720.117 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#52: RNA chain | Mass: 43923.266 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#53: RNA chain | Mass: 186482.641 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
#54: RNA chain | Mass: 132907.203 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Chlamydomonas reinhardtii (plant) |
-Non-polymers , 3 types, 71 molecules
#55: Chemical | ChemComp-ZN / | ||
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#56: Chemical | ChemComp-MG / #57: Chemical | ChemComp-FES / | |
-Details
Has ligand of interest | N |
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-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: Large subunit of the mitochondrial ribosome of Chlamydomonas reinhardtii Type: RIBOSOME / Entity ID: #1-#54 / Source: NATURAL |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Chlamydomonas reinhardtii (plant) / Organelle: Mitochondria |
Buffer solution | pH: 7.6 |
Specimen | Conc.: 1.5 mg/ml / 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 % |
-Electron microscopy imaging
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
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Microscopy | Model: FEI TALOS ARCTICA |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 500 nm |
Specimen holder | Cryogen: NITROGEN |
Image recording | Electron dose: 45 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 |
Image scans | Movie frames/image: 65 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||
3D reconstruction | Resolution: 3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 101291 / Num. of class averages: 2 / Symmetry type: POINT |