+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13112 | ||||||||||||||||||
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Title | Cryo-EM structure of the 6 hpf zebrafish embryo 80S ribosome | ||||||||||||||||||
Map data | |||||||||||||||||||
Sample |
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Function / homology | Function and homology information optokinetic behavior / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / RMTs methylate histone arginines / TNFR1-mediated ceramide production / : / : / Formation of a pool of free 40S subunits / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition ...optokinetic behavior / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / RMTs methylate histone arginines / TNFR1-mediated ceramide production / : / : / Formation of a pool of free 40S subunits / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / : / Protein methylation / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / mTORC1-mediated signalling / brain segmentation / TNFR1-induced NF-kappa-B signaling pathway / Regulation of TNFR1 signaling / convergent extension involved in gastrulation / : / primitive hemopoiesis / embryonic retina morphogenesis in camera-type eye / mitochondrial cytochrome c oxidase assembly / hemoglobin biosynthetic process / : / chordate embryonic development / embryonic brain development / positive regulation of gastrulation / ribosomal subunit / exocrine pancreas development / anatomical structure development / laminin receptor activity / mitochondrial large ribosomal subunit / positive regulation of translational fidelity / definitive hemopoiesis / pancreas development / regulation of establishment of cell polarity / cytoplasmic side of rough endoplasmic reticulum membrane / regulation of innate immune response / erythrocyte development / negative regulation of Wnt signaling pathway / regulation of cell division / erythrocyte maturation / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / mitochondrial respiratory chain complex I assembly / protein-RNA complex assembly / translation regulator activity / hemopoiesis / intrinsic apoptotic signaling pathway by p53 class mediator / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / hematopoietic stem cell differentiation / rough endoplasmic reticulum / vasculogenesis / laminin binding / rescue of stalled ribosome / maturation of SSU-rRNA / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / positive regulation of apoptotic signaling pathway / maturation of LSU-rRNA / ribosomal large subunit biogenesis / DNA-(apurinic or apyrimidinic site) lyase / small-subunit processome / erythrocyte differentiation / protein kinase C binding / brain development / modification-dependent protein catabolic process / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / regulation of erythrocyte differentiation / ribosomal large subunit assembly / small ribosomal subunit rRNA binding / protein tag activity / spindle / ribosomal small subunit assembly / rRNA processing / cytosolic small ribosomal subunit / regulation of protein localization / large ribosomal subunit rRNA binding / ribosome binding / large ribosomal subunit / glucose homeostasis / regulation of translation / T cell differentiation in thymus / cytoplasmic translation / small ribosomal subunit / 5S rRNA binding / cytosolic large ribosomal subunit / angiogenesis / mitochondrial inner membrane / negative regulation of translation / rRNA binding / protein ubiquitination / regulation of cell cycle / ribosome / structural constituent of ribosome / positive regulation of protein phosphorylation / ribonucleoprotein complex / translation Similarity search - Function | ||||||||||||||||||
Biological species | Danio rerio (zebrafish) / Zebrafish, Brachydanio rerio | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.4 Å | ||||||||||||||||||
Authors | Leesch F / Lorenzo-Orts L / Grishkovskaya I / Kandolf S / Belacic K / Meinhart A / Haselbach D / Pauli A | ||||||||||||||||||
Funding support | Austria, Switzerland, 5 items
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Citation | Journal: Nature / Year: 2023 Title: A molecular network of conserved factors keeps ribosomes dormant in the egg. Authors: Friederike Leesch / Laura Lorenzo-Orts / Carina Pribitzer / Irina Grishkovskaya / Josef Roehsner / Anastasia Chugunova / Manuel Matzinger / Elisabeth Roitinger / Katarina Belačić / Susanne ...Authors: Friederike Leesch / Laura Lorenzo-Orts / Carina Pribitzer / Irina Grishkovskaya / Josef Roehsner / Anastasia Chugunova / Manuel Matzinger / Elisabeth Roitinger / Katarina Belačić / Susanne Kandolf / Tzi-Yang Lin / Karl Mechtler / Anton Meinhart / David Haselbach / Andrea Pauli / Abstract: Ribosomes are produced in large quantities during oogenesis and are stored in the egg. However, the egg and early embryo are translationally repressed. Here, using mass spectrometry and cryo-electron ...Ribosomes are produced in large quantities during oogenesis and are stored in the egg. However, the egg and early embryo are translationally repressed. Here, using mass spectrometry and cryo-electron microscopy analyses of ribosomes isolated from zebrafish (Danio rerio) and Xenopus laevis eggs and embryos, we provide molecular evidence that ribosomes transition from a dormant state to an active state during the first hours of embryogenesis. Dormant ribosomes are associated with four conserved factors that form two modules, consisting of Habp4-eEF2 and death associated protein 1b (Dap1b) or Dap in complex with eIF5a. Both modules occupy functionally important sites and act together to stabilize ribosomes and repress translation. Dap1b (also known as Dapl1 in mammals) is a newly discovered translational inhibitor that stably inserts into the polypeptide exit tunnel. Addition of recombinant zebrafish Dap1b protein is sufficient to block translation and reconstitute the dormant egg ribosome state in a mammalian translation extract in vitro. Thus, a developmentally programmed, conserved ribosome state has a key role in ribosome storage and translational repression in the egg. | ||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_13112.map.gz | 311.3 MB | EMDB map data format | |
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Header (meta data) | emd-13112-v30.xml emd-13112.xml | 91.5 KB 91.5 KB | Display Display | EMDB header |
Images | emd_13112.png | 87.5 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13112 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13112 | HTTPS FTP |
-Related structure data
Related structure data | 7oybMC 7oyaC 7oycC 7oydC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_13112.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Voxel size | X=Y=Z: 1.06 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
+Entire : 80S ribosome from 6 hpf zebrafish embryos
+Supramolecule #1: 80S ribosome from 6 hpf zebrafish embryos
+Macromolecule #1: 18S rRNA
+Macromolecule #2: 28S rRNA
+Macromolecule #3: 5S rRNA
+Macromolecule #4: 5.8S rRNA
+Macromolecule #5: 60S ribosomal protein L8
+Macromolecule #6: 40S ribosomal protein SA
+Macromolecule #7: Ribosomal protein L3
+Macromolecule #8: 40S ribosomal protein S3a
+Macromolecule #9: Ribosomal protein L4
+Macromolecule #10: 40S ribosomal protein S2
+Macromolecule #11: Ribosomal protein L5b
+Macromolecule #12: DNA-(apurinic or apyrimidinic site) lyase
+Macromolecule #13: 60S ribosomal protein L6
+Macromolecule #14: 40S ribosomal protein S4, X isoform
+Macromolecule #15: Ribosomal protein L7
+Macromolecule #16: Ribosomal protein S5
+Macromolecule #17: 60S ribosomal protein L7a
+Macromolecule #18: 40S ribosomal protein S6
+Macromolecule #19: 60S ribosomal protein L9
+Macromolecule #20: 40S ribosomal protein S7
+Macromolecule #21: 60S ribosomal protein L10
+Macromolecule #22: 40S ribosomal protein S8
+Macromolecule #23: 60S ribosomal protein L11
+Macromolecule #24: 40S ribosomal protein S9
+Macromolecule #25: Ribosomal protein S10
+Macromolecule #26: 60S ribosomal protein L13
+Macromolecule #27: 40S ribosomal protein S11
+Macromolecule #28: 60S ribosomal protein L14
+Macromolecule #29: Ribosomal protein L15
+Macromolecule #30: 40S ribosomal protein S13
+Macromolecule #31: 60S ribosomal protein L13a
+Macromolecule #32: Ribosomal protein S14
+Macromolecule #33: 60S ribosomal protein L17
+Macromolecule #34: 40S ribosomal protein S15
+Macromolecule #35: Ribosomal protein L18
+Macromolecule #36: Ribosomal protein S16
+Macromolecule #37: 60S ribosomal protein L19
+Macromolecule #38: 40S ribosomal protein S17
+Macromolecule #39: 60S ribosomal protein L18a
+Macromolecule #40: 40S ribosomal protein S18
+Macromolecule #41: 60S ribosomal protein L21
+Macromolecule #42: 40S ribosomal protein S19
+Macromolecule #43: Ribosomal protein L22
+Macromolecule #44: 40S ribosomal protein S20
+Macromolecule #45: 60S ribosomal protein L23
+Macromolecule #46: 40S ribosomal protein S21
+Macromolecule #47: 60S ribosomal protein L24
+Macromolecule #48: 40S ribosomal protein S15a
+Macromolecule #49: Ribosomal protein L23a
+Macromolecule #50: 40S ribosomal protein S23
+Macromolecule #51: ATPase H+ transporting V0 subunit e1
+Macromolecule #52: 40S ribosomal protein S24
+Macromolecule #53: 60S ribosomal protein L27
+Macromolecule #54: 40S ribosomal protein S25
+Macromolecule #55: 60S ribosomal protein L27a
+Macromolecule #56: 40S ribosomal protein S26
+Macromolecule #57: 60S ribosomal protein L29
+Macromolecule #58: 40S ribosomal protein S27
+Macromolecule #59: 60S ribosomal protein L30
+Macromolecule #60: 40S ribosomal protein S28
+Macromolecule #61: 60S ribosomal protein L31
+Macromolecule #62: 40S ribosomal protein S29
+Macromolecule #63: Ribosomal protein L32
+Macromolecule #64: 40S ribosomal protein S30
+Macromolecule #65: 60S ribosomal protein L35a
+Macromolecule #66: 60S ribosomal protein L34
+Macromolecule #67: Guanine nucleotide-binding protein subunit beta-2-like 1
+Macromolecule #68: 60S ribosomal protein L35
+Macromolecule #69: 60S ribosomal protein L36
+Macromolecule #70: Ribosomal protein L37
+Macromolecule #71: 60S ribosomal protein L38
+Macromolecule #72: Ribosomal protein L39
+Macromolecule #73: 60S ribosomal protein L40
+Macromolecule #74: Rpl41
+Macromolecule #75: 60S ribosomal protein L36a
+Macromolecule #76: Zgc:171772
+Macromolecule #77: 60S ribosomal protein L28
+Macromolecule #78: MAGNESIUM ION
+Macromolecule #79: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.6 Component:
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Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2.0 nm / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 70 % / Instrument: LEICA EM GP |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Number grids imaged: 1 / Number real images: 11860 / Average electron dose: 48.3 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Particle selection | Number selected: 1285670 |
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Initial angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 2.14) |
Final 3D classification | Number classes: 3 / Software - Name: cryoSPARC |
Final angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 2.14) |
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 2.4 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 2.14) / Details: composite map of separately refined subunits / Number images used: 775288 |
-Atomic model buiding 1
Refinement | Protocol: OTHER |
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Output model | PDB-7oyb: |