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Yorodumi- EMDB-12870: Structure of a human mitochondrial ribosome large subunit assembl... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12870 | ||||||||||||||||||
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Title | Structure of a human mitochondrial ribosome large subunit assembly intermediate in complex with MTERF4-NSUN4 and GTPBP5 (dataset2). | ||||||||||||||||||
Map data | Global refinement map. | ||||||||||||||||||
Sample |
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Keywords | Mitochondria / Ribosome / Biogenesis / GTPase / NSUN4 / MTERF4 | ||||||||||||||||||
Function / homology | Function and homology information regulation of respiratory system process / tRNA (cytidine-5-)-methyltransferase activity / rRNA modification in the mitochondrion / regulation of mitochondrial translation / negative regulation of mitochondrial translation / mitochondrial large ribosomal subunit assembly / rRNA (cytosine-C5-)-methyltransferase activity / Complex I biogenesis / protein lipoylation / negative regulation of ribosome biogenesis ...regulation of respiratory system process / tRNA (cytidine-5-)-methyltransferase activity / rRNA modification in the mitochondrion / regulation of mitochondrial translation / negative regulation of mitochondrial translation / mitochondrial large ribosomal subunit assembly / rRNA (cytosine-C5-)-methyltransferase activity / Complex I biogenesis / protein lipoylation / negative regulation of ribosome biogenesis / Mitochondrial Fatty Acid Beta-Oxidation / Protein lipoylation / rRNA import into mitochondrion / mitochondrial [2Fe-2S] assembly complex / Respiratory electron transport / rRNA methyltransferase activity / mitochondrial transcription / mitochondrial translational termination / mitochondrial translational elongation / mitochondrial ribosome assembly / translation release factor activity, codon nonspecific / positive regulation of mitochondrial translation / microprocessor complex / Mitochondrial translation elongation / Mitochondrial translation termination / Mitochondrial translation initiation / protein targeting to mitochondrion / camera-type eye development / iron-sulfur cluster assembly complex / mitochondrial large ribosomal subunit / mitochondrial fission / mitochondrial large ribosomal subunit binding / Hydrolases; Acting on ester bonds; Endoribonucleases producing 5'-phosphomonoesters / peptidyl-tRNA hydrolase / mitochondrial ribosome / mitochondrial small ribosomal subunit / rRNA methylation / mitochondrial translation / aminoacyl-tRNA hydrolase activity / [2Fe-2S] cluster assembly / iron-sulfur cluster assembly / ribosomal large subunit binding / proton motive force-driven mitochondrial ATP synthesis / respiratory chain complex I / : / mitochondrial electron transport, NADH to ubiquinone / mitochondrial respiratory chain complex I assembly / acyl binding / anatomical structure morphogenesis / acyl carrier activity / RNA processing / Mitochondrial protein degradation / aerobic respiration / rescue of stalled ribosome / ribosomal large subunit biogenesis / Transferases; Transferring one-carbon groups; Methyltransferases / cellular response to leukemia inhibitory factor / methyltransferase activity / fatty acid binding / mitochondrial membrane / fibrillar center / fatty acid biosynthetic process / rRNA processing / double-stranded RNA binding / small ribosomal subunit rRNA binding / cell junction / ribosome biogenesis / heart development / 5S rRNA binding / large ribosomal subunit rRNA binding / double-stranded DNA binding / endonuclease activity / mitochondrial inner membrane / negative regulation of translation / nuclear body / rRNA binding / ribosome / structural constituent of ribosome / mitochondrial matrix / ribonucleoprotein complex / translation / protein domain specific binding / GTPase activity / mRNA binding / nucleotide binding / calcium ion binding / synapse / regulation of DNA-templated transcription / GTP binding / nucleolus / apoptotic process / magnesium ion binding / mitochondrion / RNA binding / extracellular space / nucleoplasm / nucleus / plasma membrane / cytosol Similarity search - Function | ||||||||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | ||||||||||||||||||
Authors | Hillen HS / Lavdovskaia E | ||||||||||||||||||
Funding support | Germany, 5 items
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Citation | Journal: Nat Commun / Year: 2021 Title: Structural basis of GTPase-mediated mitochondrial ribosome biogenesis and recycling. Authors: Hauke S Hillen / Elena Lavdovskaia / Franziska Nadler / Elisa Hanitsch / Andreas Linden / Katherine E Bohnsack / Henning Urlaub / Ricarda Richter-Dennerlein / Abstract: Ribosome biogenesis requires auxiliary factors to promote folding and assembly of ribosomal proteins and RNA. Particularly, maturation of the peptidyl transferase center (PTC) is mediated by ...Ribosome biogenesis requires auxiliary factors to promote folding and assembly of ribosomal proteins and RNA. Particularly, maturation of the peptidyl transferase center (PTC) is mediated by conserved GTPases, but the molecular basis is poorly understood. Here, we define the mechanism of GTPase-driven maturation of the human mitochondrial large ribosomal subunit (mtLSU) using endogenous complex purification, in vitro reconstitution and cryo-EM. Structures of transient native mtLSU assembly intermediates that accumulate in GTPBP6-deficient cells reveal how the biogenesis factors GTPBP5, MTERF4 and NSUN4 facilitate PTC folding. Addition of recombinant GTPBP6 reconstitutes late mtLSU biogenesis in vitro and shows that GTPBP6 triggers a molecular switch and progression to a near-mature PTC state. Additionally, cryo-EM analysis of GTPBP6-treated mature mitochondrial ribosomes reveals the structural basis for the dual-role of GTPBP6 in ribosome biogenesis and recycling. Together, these results provide a framework for understanding step-wise PTC folding as a critical conserved quality control checkpoint. #1: Journal: Biorxiv / Year: 2021 Title: Structural basis of GTPase-mediated mitochondrial ribosome biogenesis and recycling Authors: Hillen HS / Lavdovskaia E / Nadler F / Hanitsch E / Linden A / Bohnsack KE / Urlaub H / Richter-Dennerlein R | ||||||||||||||||||
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_12870.map.gz | 112.9 MB | EMDB map data format | |
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Header (meta data) | emd-12870-v30.xml emd-12870.xml | 88.9 KB 88.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_12870_fsc.xml | 12.4 KB | Display | FSC data file |
Images | emd_12870.png | 110.9 KB | ||
Masks | emd_12870_msk_1.map emd_12870_msk_2.map | 163.6 MB 163.6 MB | Mask map | |
Filedesc metadata | emd-12870.cif.gz | 17.4 KB | ||
Others | emd_12870_additional_1.map.gz emd_12870_additional_2.map.gz emd_12870_additional_3.map.gz emd_12870_additional_4.map.gz emd_12870_half_map_1.map.gz emd_12870_half_map_2.map.gz | 16.3 MB 110.7 MB 111.8 MB 111.8 MB 113.5 MB 113.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12870 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12870 | HTTPS FTP |
-Validation report
Summary document | emd_12870_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_12870_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_12870_validation.xml.gz | 19.7 KB | Display | |
Data in CIF | emd_12870_validation.cif.gz | 26.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12870 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12870 | HTTPS FTP |
-Related structure data
Related structure data | 7of5MC 7of0C 7of2C 7of3C 7of4C 7of6C 7of7C 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_12870.map.gz / Format: CCP4 / Size: 163.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Global refinement map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.05 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_12870_msk_1.map | ||||||||||||
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-Mask #2
File | emd_12870_msk_2.map | ||||||||||||
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-Additional map: Global post-processed map.
File | emd_12870_additional_1.map | ||||||||||||
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Annotation | Global post-processed map. | ||||||||||||
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-Additional map: Focused refinement map.
File | emd_12870_additional_2.map | ||||||||||||
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Annotation | Focused refinement map. | ||||||||||||
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-Additional map: Focused refinement half-map 1.
File | emd_12870_additional_3.map | ||||||||||||
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Annotation | Focused refinement half-map 1. | ||||||||||||
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-Additional map: Focused refinement half-map 2.
File | emd_12870_additional_4.map | ||||||||||||
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Annotation | Focused refinement half-map 2. | ||||||||||||
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Density Histograms |
-Half map: Global refinement half-map 1.
File | emd_12870_half_map_1.map | ||||||||||||
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Annotation | Global refinement half-map 1. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Global refinement half-map 2.
File | emd_12870_half_map_2.map | ||||||||||||
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Annotation | Global refinement half-map 2. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Human mitochondrial ribosome large subunit assembly intermediate ...
+Supramolecule #1: Human mitochondrial ribosome large subunit assembly intermediate ...
+Macromolecule #1: 39S ribosomal protein L32, mitochondrial
+Macromolecule #2: 39S ribosomal protein L33, mitochondrial
+Macromolecule #3: 39S ribosomal protein L34, mitochondrial
+Macromolecule #4: 39S ribosomal protein L35, mitochondrial
+Macromolecule #5: 39S ribosomal protein L36, mitochondrial
+Macromolecule #6: 39S ribosomal protein L37, mitochondrial
+Macromolecule #7: 39S ribosomal protein L38, mitochondrial
+Macromolecule #8: 39S ribosomal protein L39, mitochondrial
+Macromolecule #9: 39S ribosomal protein L40, mitochondrial
+Macromolecule #10: 39S ribosomal protein L41, mitochondrial
+Macromolecule #13: 5-methylcytosine rRNA methyltransferase NSUN4
+Macromolecule #14: 39S ribosomal protein L2, mitochondrial
+Macromolecule #15: 39S ribosomal protein L3, mitochondrial
+Macromolecule #16: 39S ribosomal protein L4, mitochondrial
+Macromolecule #17: Transcription termination factor 4, mitochondrial
+Macromolecule #18: 39S ribosomal protein L9, mitochondrial
+Macromolecule #19: 39S ribosomal protein L10, mitochondrial
+Macromolecule #20: 39S ribosomal protein L11, mitochondrial
+Macromolecule #21: 39S ribosomal protein L13, mitochondrial
+Macromolecule #22: 39S ribosomal protein L14, mitochondrial
+Macromolecule #23: 39S ribosomal protein L15, mitochondrial
+Macromolecule #24: 39S ribosomal protein L16, mitochondrial
+Macromolecule #25: 39S ribosomal protein L17, mitochondrial
+Macromolecule #26: 39S ribosomal protein L18, mitochondrial
+Macromolecule #27: 39S ribosomal protein L19, mitochondrial
+Macromolecule #28: 39S ribosomal protein L20, mitochondrial
+Macromolecule #29: 39S ribosomal protein L21, mitochondrial
+Macromolecule #30: 39S ribosomal protein L22, mitochondrial
+Macromolecule #31: 39S ribosomal protein L23, mitochondrial
+Macromolecule #32: 39S ribosomal protein L24, mitochondrial
+Macromolecule #33: 39S ribosomal protein L27, mitochondrial
+Macromolecule #34: 39S ribosomal protein L28, mitochondrial
+Macromolecule #35: 39S ribosomal protein L47, mitochondrial
+Macromolecule #36: 39S ribosomal protein L30, mitochondrial
+Macromolecule #37: 39S ribosomal protein L42, mitochondrial
+Macromolecule #38: 39S ribosomal protein L43, mitochondrial
+Macromolecule #39: 39S ribosomal protein L44, mitochondrial
+Macromolecule #40: 39S ribosomal protein L45, mitochondrial
+Macromolecule #41: 39S ribosomal protein L46, mitochondrial
+Macromolecule #42: 39S ribosomal protein L48, mitochondrial
+Macromolecule #43: 39S ribosomal protein L49, mitochondrial
+Macromolecule #44: 39S ribosomal protein L50, mitochondrial
+Macromolecule #45: 39S ribosomal protein L51, mitochondrial
+Macromolecule #46: 39S ribosomal protein L52, mitochondrial
+Macromolecule #47: 39S ribosomal protein L53, mitochondrial
+Macromolecule #48: 39S ribosomal protein L55, mitochondrial
+Macromolecule #49: Ribosomal protein 63, mitochondrial
+Macromolecule #50: Peptidyl-tRNA hydrolase ICT1, mitochondrial
+Macromolecule #51: Growth arrest and DNA damage-inducible proteins-interacting protein 1
+Macromolecule #52: 39S ribosomal protein S18a, mitochondrial
+Macromolecule #53: 39S ribosomal protein S30, mitochondrial
+Macromolecule #54: Mitochondrial assembly of ribosomal large subunit protein 1
+Macromolecule #55: MIEF1 upstream open reading frame protein
+Macromolecule #56: Acyl carrier protein, mitochondrial
+Macromolecule #57: Mitochondrial ribosome-associated GTPase 2
+Macromolecule #11: 16S ribosomal RNA
+Macromolecule #12: Mitochondrial tRNAVal
+Macromolecule #58: ZINC ION
+Macromolecule #59: MAGNESIUM ION
+Macromolecule #60: GUANOSINE-5'-TRIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil R3.5/1 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 0.25 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 37.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated defocus max: 2.1 µm / Calibrated defocus min: 0.3 µm / Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |