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Yorodumi- EMDB-10021: Structure of human mitochondrial 28S ribosome in complex with mit... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-10021 | |||||||||
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Title | Structure of human mitochondrial 28S ribosome in complex with mitochondrial IF3 | |||||||||
Map data | Composite map, local resolution filtered | |||||||||
Sample |
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Keywords | ribosomal small subunit / initiation complex / initiation factor 3 / mitochondria / RIBOSOME | |||||||||
Function / homology | Function and homology information mitochondrial translational initiation / mitochondrial ribosome binding / translation factor activity, RNA binding / mitochondrial ribosome assembly / ribosome disassembly / positive regulation of mitochondrial translation / Mitochondrial translation elongation / Mitochondrial translation termination / Mitochondrial translation initiation / negative regulation of mitotic nuclear division ...mitochondrial translational initiation / mitochondrial ribosome binding / translation factor activity, RNA binding / mitochondrial ribosome assembly / ribosome disassembly / positive regulation of mitochondrial translation / Mitochondrial translation elongation / Mitochondrial translation termination / Mitochondrial translation initiation / negative regulation of mitotic nuclear division / mitochondrial ribosome / mitochondrial small ribosomal subunit / mitochondrial translation / positive regulation of proteolysis / ribosomal small subunit binding / Mitochondrial protein degradation / translation initiation factor activity / apoptotic signaling pathway / fibrillar center / small ribosomal subunit rRNA binding / cell junction / ribosome binding / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / nuclear membrane / cell population proliferation / tRNA binding / mitochondrial inner membrane / rRNA binding / ribosome / structural constituent of ribosome / mitochondrial matrix / translation / protein domain specific binding / intracellular membrane-bounded organelle / mRNA binding / GTP binding / nucleolus / mitochondrion / RNA binding / nucleoplasm / nucleus / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.97 Å | |||||||||
Authors | Itoh Y / Khawaja A | |||||||||
Citation | Journal: Nat Commun / Year: 2020 Title: Distinct pre-initiation steps in human mitochondrial translation. Authors: Anas Khawaja / Yuzuru Itoh / Cristina Remes / Henrik Spåhr / Olessya Yukhnovets / Henning Höfig / Alexey Amunts / Joanna Rorbach / Abstract: Translation initiation in human mitochondria relies upon specialized mitoribosomes and initiation factors, mtIF2 and mtIF3, which have diverged from their bacterial counterparts. Here we report two ...Translation initiation in human mitochondria relies upon specialized mitoribosomes and initiation factors, mtIF2 and mtIF3, which have diverged from their bacterial counterparts. Here we report two distinct mitochondrial pre-initiation assembly steps involving those factors. Single-particle cryo-EM revealed that in the first step, interactions between mitochondria-specific protein mS37 and mtIF3 keep the small mitoribosomal subunit in a conformation favorable for a subsequent accommodation of mtIF2 in the second step. Combination with fluorescence cross-correlation spectroscopy analyses suggests that mtIF3 promotes complex assembly without mRNA or initiator tRNA binding, where exclusion is achieved by the N-terminal and C-terminal domains of mtIF3. Finally, the association of large mitoribosomal subunit is required for initiator tRNA and leaderless mRNA recruitment to form a stable initiation complex. These data reveal fundamental aspects of mammalian protein synthesis that are specific to mitochondria. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_10021.map.gz | 14.1 MB | EMDB map data format | |
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Header (meta data) | emd-10021-v30.xml emd-10021.xml | 64.1 KB 64.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_10021_fsc.xml | 17.1 KB | Display | FSC data file |
Images | emd_10021.png | 121 KB | ||
Masks | emd_10021_msk_1.map | 421.9 MB | Mask map | |
Filedesc metadata | emd-10021.cif.gz | 14.1 KB | ||
Others | emd_10021_additional.map.gz emd_10021_half_map_1.map.gz emd_10021_half_map_2.map.gz | 387.3 MB 340.5 MB 340.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-10021 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10021 | HTTPS FTP |
-Validation report
Summary document | emd_10021_validation.pdf.gz | 747.7 KB | Display | EMDB validaton report |
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Full document | emd_10021_full_validation.pdf.gz | 747.3 KB | Display | |
Data in XML | emd_10021_validation.xml.gz | 23.8 KB | Display | |
Data in CIF | emd_10021_validation.cif.gz | 31.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10021 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10021 | HTTPS FTP |
-Related structure data
Related structure data | 6rw4MC 6rw5C M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_10021.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Composite map, local resolution filtered | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.83 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_10021_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: Overall map without sharpening
File | emd_10021_additional.map | ||||||||||||
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Annotation | Overall map without sharpening | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_10021_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_10021_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : IF3 bound mitochondrial translation pre-initiation complex
+Supramolecule #1: IF3 bound mitochondrial translation pre-initiation complex
+Supramolecule #2: Ribosome
+Supramolecule #3: IF3
+Macromolecule #1: 12S mitochondrial rRNA
+Macromolecule #2: 28S ribosomal protein S2, mitochondrial
+Macromolecule #3: 28S ribosomal protein S24, mitochondrial
+Macromolecule #4: 28S ribosomal protein S5, mitochondrial
+Macromolecule #5: 28S ribosomal protein S6, mitochondrial
+Macromolecule #6: 28S ribosomal protein S7, mitochondrial
+Macromolecule #7: 28S ribosomal protein S9, mitochondrial
+Macromolecule #8: 28S ribosomal protein S10, mitochondrial
+Macromolecule #9: 28S ribosomal protein S11, mitochondrial
+Macromolecule #10: 28S ribosomal protein S12, mitochondrial
+Macromolecule #11: 28S ribosomal protein S14, mitochondrial
+Macromolecule #12: 28S ribosomal protein S15, mitochondrial
+Macromolecule #13: 28S ribosomal protein S16, mitochondrial
+Macromolecule #14: 28S ribosomal protein S17, mitochondrial
+Macromolecule #15: 28S ribosomal protein S18b, mitochondrial
+Macromolecule #16: 28S ribosomal protein S18c, mitochondrial
+Macromolecule #17: 28S ribosomal protein S21, mitochondrial
+Macromolecule #18: 28S ribosomal protein S22, mitochondrial
+Macromolecule #19: 28S ribosomal protein S23, mitochondrial
+Macromolecule #20: 28S ribosomal protein S25, mitochondrial
+Macromolecule #21: 28S ribosomal protein S26, mitochondrial
+Macromolecule #22: 28S ribosomal protein S27, mitochondrial
+Macromolecule #23: 28S ribosomal protein S28, mitochondrial
+Macromolecule #24: 28S ribosomal protein S29, mitochondrial
+Macromolecule #25: 28S ribosomal protein S31, mitochondrial
+Macromolecule #26: 28S ribosomal protein S33, mitochondrial
+Macromolecule #27: 28S ribosomal protein S34, mitochondrial
+Macromolecule #28: 28S ribosomal protein S35, mitochondrial
+Macromolecule #29: Coiled-coil-helix-coiled-coil-helix domain-containing protein 1
+Macromolecule #30: Aurora kinase A-interacting protein
+Macromolecule #31: Pentatricopeptide repeat domain-containing protein 3, mitochondrial
+Macromolecule #32: Translation initiation factor IF-3, mitochondrial
+Macromolecule #33: NICOTINAMIDE-ADENINE-DINUCLEOTIDE
+Macromolecule #34: SPERMINE
+Macromolecule #35: STREPTOMYCIN
+Macromolecule #36: MAGNESIUM ION
+Macromolecule #37: POTASSIUM ION
+Macromolecule #38: ZINC ION
+Macromolecule #39: FE2/S2 (INORGANIC) CLUSTER
+Macromolecule #40: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #41: PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER
+Macromolecule #42: water
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.7 mg/mL | ||||||||||||||||||
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Buffer | pH: 7.5 Component:
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Grid | Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. Details: The grid was coated with 3 nm carbon film prior to use | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 2-20 / Number grids imaged: 1 / Number real images: 13831 / Average exposure time: 4.0 sec. / Average electron dose: 1.5 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Calibrated defocus max: 5.0 µm / Calibrated defocus min: 0.25 µm / Calibrated magnification: 165000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.2 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
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Refinement | Space: REAL / Protocol: AB INITIO MODEL / Overall B value: 36 / Target criteria: Cross-correlation coefficient |
Output model | PDB-6rw4: |