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Yorodumi- EMDB-3461: Cryo-EM structure of a native ribosome-Ski2-Ski3-Ski8 complex fro... -
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Basic information
| Entry | Database: EMDB / ID: EMD-3461 | |||||||||
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| Title | Cryo-EM structure of a native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae | |||||||||
Map data | Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae | |||||||||
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Keywords | cryo-EM / ribosome / RNA / helicase | |||||||||
| Function / homology | Function and homology informationcytoplasmic translational elongation through polyproline stretches / Hypusine synthesis from eIF5A-lysine / CAT tailing / translational frameshifting / mRNA decay by 3' to 5' exoribonuclease / protein-DNA complex assembly / cytoplasmic translational elongation / Ski complex / nuclear-transcribed mRNA catabolic process, 3'-5' exonucleolytic nonsense-mediated decay / cytoplasmic translational termination ...cytoplasmic translational elongation through polyproline stretches / Hypusine synthesis from eIF5A-lysine / CAT tailing / translational frameshifting / mRNA decay by 3' to 5' exoribonuclease / protein-DNA complex assembly / cytoplasmic translational elongation / Ski complex / nuclear-transcribed mRNA catabolic process, 3'-5' exonucleolytic nonsense-mediated decay / cytoplasmic translational termination / positive regulation of translational elongation / nuclear-transcribed mRNA catabolic process, non-stop decay / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, LSU-rRNA,5S) / regulation of amino acid metabolic process / negative regulation of glucose mediated signaling pathway / positive regulation of translational fidelity / RMTs methylate histone arginines / Protein methylation / response to cycloheximide / translational readthrough / mTORC1-mediated signalling / Protein hydroxylation / reciprocal meiotic recombination / pre-mRNA 5'-splice site binding / GDP-dissociation inhibitor activity / Ribosome Quality Control (RQC) complex extracts and degrades nascent peptide / ascospore wall assembly / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / cytosolic large ribosomal subunit assembly / Ribosomal scanning and start codon recognition / nonfunctional rRNA decay / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / preribosome, small subunit precursor / mRNA destabilization / cytoplasmic translational initiation / Major pathway of rRNA processing in the nucleolus and cytosol / nuclear chromosome / PELO:HBS1L and ABCE1 dissociate a ribosome on a non-stop mRNA / mRNA catabolic process / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / protein-RNA complex assembly / Formation of a pool of free 40S subunits / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / preribosome, large subunit precursor / positive regulation of protein kinase activity / L13a-mediated translational silencing of Ceruloplasmin expression / negative regulation of mRNA splicing, via spliceosome / negative regulation of translational frameshifting / ribosomal large subunit export from nucleus / translational elongation / nuclear-transcribed mRNA catabolic process / translation elongation factor activity / G-protein alpha-subunit binding / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / 90S preribosome / ribosomal subunit export from nucleus / translational termination / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / ribosome-associated ubiquitin-dependent protein catabolic process / maturation of LSU-rRNA / 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) / translation regulator activity / DNA-(apurinic or apyrimidinic site) endonuclease activity / ribosomal small subunit export from nucleus / translation initiation factor activity / cellular response to amino acid starvation / protein kinase C binding / macroautophagy / rescue of stalled cytosolic ribosome / ribosome assembly / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / translational initiation / maintenance of translational fidelity / modification-dependent protein catabolic process / cytoplasmic stress granule / protein tag activity / rRNA processing / cytosolic ribosome / ribosomal small subunit assembly / ribosome biogenesis / ribosome binding / protein-containing complex assembly / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / 5S rRNA binding / ribosomal large subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / RNA helicase activity Similarity search - Function | |||||||||
| Biological species | ![]() ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||
Authors | Schmidt C / Kowalinski E | |||||||||
Citation | Journal: Science / Year: 2016Title: The cryo-EM structure of a ribosome-Ski2-Ski3-Ski8 helicase complex. Authors: Christian Schmidt / Eva Kowalinski / Vivekanandan Shanmuganathan / Quentin Defenouillère / Katharina Braunger / André Heuer / Markus Pech / Abdelkader Namane / Otto Berninghausen / ...Authors: Christian Schmidt / Eva Kowalinski / Vivekanandan Shanmuganathan / Quentin Defenouillère / Katharina Braunger / André Heuer / Markus Pech / Abdelkader Namane / Otto Berninghausen / Micheline Fromont-Racine / Alain Jacquier / Elena Conti / Thomas Becker / Roland Beckmann / ![]() Abstract: Ski2-Ski3-Ski8 (Ski) is a helicase complex functioning with the RNA-degrading exosome to mediate the 3'-5' messenger RNA (mRNA) decay in turnover and quality-control pathways. We report that the Ski ...Ski2-Ski3-Ski8 (Ski) is a helicase complex functioning with the RNA-degrading exosome to mediate the 3'-5' messenger RNA (mRNA) decay in turnover and quality-control pathways. We report that the Ski complex directly associates with 80S ribosomes presenting a short mRNA 3' overhang. We determined the structure of an endogenous ribosome-Ski complex using cryo-electron microscopy (EM) with a local resolution of the Ski complex ranging from 4 angstroms (Å) in the core to about 10 Å for intrinsically flexible regions. Ribosome binding displaces the autoinhibitory domain of the Ski2 helicase, positioning it in an open conformation near the ribosomal mRNA entry tunnel. We observe that the mRNA 3' overhang is threaded directly from the small ribosomal subunit to the helicase channel of Ski2, primed for ongoing exosome-mediated 3'-5' degradation. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_3461.map.gz | 381.9 MB | EMDB map data format | |
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| Header (meta data) | emd-3461-v30.xml emd-3461.xml | 113.5 KB 113.5 KB | Display Display | EMDB header |
| Images | emd_3461.png | 38.7 KB | ||
| Filedesc metadata | emd-3461.cif.gz | 22.1 KB | ||
| Archive directory | https://data.pdbj.org/pub/emdb/structures/EMD-3461 ftp://data.pdbj.org/pub/emdb/structures/EMD-3461 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 5mc6MC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_3461.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.084 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae
+Supramolecule #1: Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae
+Macromolecule #1: 18S ribosomal RNA
+Macromolecule #35: mRNA
+Macromolecule #36: A-site tRNA
+Macromolecule #37: P-site tRNA
+Macromolecule #83: 25S ribosomal RNA
+Macromolecule #84: 5S ribosomal RNA
+Macromolecule #85: 5.8S ribosomal RNA
+Macromolecule #2: 40S ribosomal protein S3
+Macromolecule #3: 40S ribosomal protein S5
+Macromolecule #4: 40S ribosomal protein S10-A
+Macromolecule #5: 40S ribosomal protein S12
+Macromolecule #6: 40S ribosomal protein S15
+Macromolecule #7: 40S ribosomal protein S16-A
+Macromolecule #8: 40S ribosomal protein S17-B
+Macromolecule #9: 40S ribosomal protein S18-A
+Macromolecule #10: 40S ribosomal protein S19-A
+Macromolecule #11: 40S ribosomal protein S20
+Macromolecule #12: 40S ribosomal protein S25-A
+Macromolecule #13: 40S ribosomal protein S28-B
+Macromolecule #14: 40S ribosomal protein S29-A
+Macromolecule #15: Ubiquitin-40S ribosomal protein S31
+Macromolecule #16: Guanine nucleotide-binding protein subunit beta-like protein
+Macromolecule #17: 40S ribosomal protein S0-A
+Macromolecule #18: 40S ribosomal protein S1-A
+Macromolecule #19: 40S ribosomal protein S2
+Macromolecule #20: 40S ribosomal protein S4-A
+Macromolecule #21: 40S ribosomal protein S6-A
+Macromolecule #22: 40S ribosomal protein S7-A
+Macromolecule #23: 40S ribosomal protein S8-A
+Macromolecule #24: 40S ribosomal protein S9-A
+Macromolecule #25: 40S ribosomal protein S11-A
+Macromolecule #26: 40S ribosomal protein S13
+Macromolecule #27: 40S ribosomal protein S14-A
+Macromolecule #28: 40S ribosomal protein S21-A
+Macromolecule #29: 40S ribosomal protein S22-A
+Macromolecule #30: 40S ribosomal protein S23-A
+Macromolecule #31: 40S ribosomal protein S24-A
+Macromolecule #32: 40S ribosomal protein S26-A
+Macromolecule #33: 40S ribosomal protein S27-A
+Macromolecule #34: 40S ribosomal protein S30-A
+Macromolecule #38: Antiviral helicase SKI2
+Macromolecule #39: Superkiller protein 3
+Macromolecule #40: Antiviral protein SKI8
+Macromolecule #41: 60S ribosomal protein L8-A
+Macromolecule #42: 60S ribosomal protein L23-A
+Macromolecule #43: 60S ribosomal protein L36-A
+Macromolecule #44: 60S ribosomal protein L9-A
+Macromolecule #45: 60S ribosomal protein L24-A
+Macromolecule #46: 60S ribosomal protein L37-A
+Macromolecule #47: 60S ribosomal protein L11-A
+Macromolecule #48: 60S ribosomal protein L25
+Macromolecule #49: 60S ribosomal protein L38
+Macromolecule #50: 60S ribosomal protein L13-A
+Macromolecule #51: 60S ribosomal protein L26-A
+Macromolecule #52: 60S ribosomal protein L39
+Macromolecule #53: 60S ribosomal protein L14-A
+Macromolecule #54: 60S ribosomal protein L27-A
+Macromolecule #55: Ubiquitin-60S ribosomal protein L40
+Macromolecule #56: 60S ribosomal protein L42-A
+Macromolecule #57: 60S ribosomal protein L15-A
+Macromolecule #58: 60S ribosomal protein L28
+Macromolecule #59: 60S ribosomal protein L41-A
+Macromolecule #60: 60S ribosomal protein L43-A
+Macromolecule #61: 60S ribosomal protein L16-A
+Macromolecule #62: 60S ribosomal protein L29
+Macromolecule #63: 60S ribosomal protein L2-A
+Macromolecule #64: 60S ribosomal protein L17-A
+Macromolecule #65: 60S ribosomal protein L30
+Macromolecule #66: uL1
+Macromolecule #67: 60S ribosomal protein L3
+Macromolecule #68: 60S ribosomal protein L18-A
+Macromolecule #69: 60S ribosomal protein L31-A
+Macromolecule #70: 60S ribosomal protein L10
+Macromolecule #71: 60S ribosomal protein L4-A
+Macromolecule #72: 60S ribosomal protein L19-A
+Macromolecule #73: 60S ribosomal protein L32
+Macromolecule #74: 60S ribosomal protein L20-A
+Macromolecule #75: 60S ribosomal protein L5
+Macromolecule #76: 60S ribosomal protein L21-A
+Macromolecule #77: 60S ribosomal protein L33-A
+Macromolecule #78: 60S ribosomal protein L22-A
+Macromolecule #79: 60S ribosomal protein L6-A
+Macromolecule #80: 60S ribosomal protein L34-A
+Macromolecule #81: 60S ribosomal protein L7-A
+Macromolecule #82: 60S ribosomal protein L35-A
+Macromolecule #86: Eukaryotic translation initiation factor 5A-1
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.4 Component:
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| Grid | Model: Quantifoil R3/3 / Material: COPPER/PALLADIUM / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. | |||||||||||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Digitization - Frames/image: 1-10 / Number grids imaged: 2 / Average electron dose: 2.8 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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