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- EMDB-0949: Cryo-EM structure of 90S small subunit preribosomes in transition... -
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Basic information
Entry | Database: EMDB / ID: EMD-0949 | ||||||||||||||||||
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Title | Cryo-EM structure of 90S small subunit preribosomes in transition states (State A) | ||||||||||||||||||
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![]() | ribosome assembly / 90S to pre-40S transition / cryo-EM / Dhr1 / RIBOSOME | ||||||||||||||||||
Function / homology | ![]() rRNA acetylation involved in maturation of SSU-rRNA / 18S rRNA cytidine N-acetyltransferase activity / tRNA N4-acetyltransferase activity / regulation of ribosomal protein gene transcription by RNA polymerase II / rRNA small subunit pseudouridine methyltransferase Nep1 / tRNA acetylation / Noc4p-Nop14p complex / RNA fragment catabolic process / rRNA 2'-O-methylation / CURI complex ...rRNA acetylation involved in maturation of SSU-rRNA / 18S rRNA cytidine N-acetyltransferase activity / tRNA N4-acetyltransferase activity / regulation of ribosomal protein gene transcription by RNA polymerase II / rRNA small subunit pseudouridine methyltransferase Nep1 / tRNA acetylation / Noc4p-Nop14p complex / RNA fragment catabolic process / rRNA 2'-O-methylation / CURI complex / UTP-C complex / t-UTP complex / nuclear microtubule / Mpp10 complex / snoRNA guided rRNA 2'-O-methylation / Pwp2p-containing subcomplex of 90S preribosome / rRNA modification / rRNA (pseudouridine) methyltransferase activity / regulation of rRNA processing / septum digestion after cytokinesis / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / snRNA binding / tRNA re-export from nucleus / box C/D sno(s)RNA 3'-end processing / regulation of transcription by RNA polymerase I / rDNA heterochromatin / rRNA methyltransferase activity / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / box C/D methylation guide snoRNP complex / positive regulation of rRNA processing / single-stranded telomeric DNA binding / U4/U6 snRNP / tRNA export from nucleus / rRNA primary transcript binding / rRNA base methylation / Negative regulators of DDX58/IFIH1 signaling / sno(s)RNA-containing ribonucleoprotein complex / SUMOylation of RNA binding proteins / protein localization to nucleolus / U4 snRNA binding / O-methyltransferase activity / small nuclear ribonucleoprotein complex / rRNA methylation / histone H2A Q104 methyltransferase activity / mTORC1-mediated signalling / Protein hydroxylation / mRNA modification / U4 snRNP / U3 snoRNA binding / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / Formation of the ternary complex, and subsequently, the 43S complex / poly(A)+ mRNA export from nucleus / Translation initiation complex formation / preribosome, small subunit precursor / Ribosomal scanning and start codon recognition / snoRNA binding / precatalytic spliceosome / establishment of cell polarity / Major pathway of rRNA processing in the nucleolus and cytosol / positive regulation of transcription by RNA polymerase I / spliceosomal complex assembly / nucleolar large rRNA transcription by RNA polymerase I / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Formation of a pool of free 40S subunits / L13a-mediated translational silencing of Ceruloplasmin expression / 90S preribosome / Ub-specific processing proteases / 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) / regulation of translational fidelity / RNA processing / ribosomal subunit export from nucleus / U4/U6 x U5 tri-snRNP complex / vesicle-mediated transport / Transferases; Acyltransferases; Transferring groups other than aminoacyl groups / RNA endonuclease activity / enzyme activator activity / nuclear periphery / Transferases; Transferring one-carbon groups; Methyltransferases / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / translational initiation / spliceosomal complex / maintenance of translational fidelity / mRNA splicing, via spliceosome / rRNA processing / protein transport / peroxisome / ribosome biogenesis / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / cytoplasmic translation / tRNA binding Similarity search - Function | ||||||||||||||||||
Biological species | ![]() ![]() ![]() ![]() | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||||||||||||||
![]() | Du Y / Ye K | ||||||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structure of 90 small ribosomal subunit precursors in transition states. Authors: Yifei Du / Weidong An / Xing Zhu / Qi Sun / Jia Qi / Keqiong Ye / ![]() Abstract: The 90 preribosome is a large, early assembly intermediate of small ribosomal subunits that undergoes structural changes to give a pre-40 ribosome. Here, we gained insight into this transition by ...The 90 preribosome is a large, early assembly intermediate of small ribosomal subunits that undergoes structural changes to give a pre-40 ribosome. Here, we gained insight into this transition by determining cryo-electron microscopy structures of intermediates in the path from the 90 to the pre-40 The full transition is blocked by deletion of RNA helicase Dhr1. A series of structural snapshots revealed that the excised 5' external transcribed spacer (5' ETS) is degraded within 90, driving stepwise disassembly of assembly factors and ribosome maturation. The nuclear exosome, an RNA degradation machine, docks on the 90 through helicase Mtr4 and is primed to digest the 3' end of the 5' ETS. The structures resolved between 3.2- and 8.6-angstrom resolution reveal key intermediates and the critical role of 5' ETS degradation in 90 progression. | ||||||||||||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 34.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 120.2 KB 120.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 15.6 KB | Display | ![]() |
Images | ![]() | 72.9 KB | ||
Filedesc metadata | ![]() | 30.1 KB | ||
Others | ![]() ![]() | 7.6 MB 4.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 459.1 KB | Display | ![]() |
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Full document | ![]() | 458.6 KB | Display | |
Data in XML | ![]() | 14.7 KB | Display | |
Data in CIF | ![]() | 20.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6lqpMC ![]() 0950C ![]() 0951C ![]() 0952C ![]() 0953C ![]() 0954C ![]() 0955C ![]() 6lqqC ![]() 6lqrC ![]() 6lqsC ![]() 6lqtC ![]() 6lquC ![]() 6lqvC C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.334 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: 5' domain map
File | emd_0949_additional_1.map | ||||||||||||
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Annotation | 5' domain map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Utp20 C map
File | emd_0949_additional_2.map | ||||||||||||
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Annotation | Utp20 C map | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
+Entire : 90S pre-ribosome (Dhr1-depleted, state A)
+Supramolecule #1: 90S pre-ribosome (Dhr1-depleted, state A)
+Macromolecule #1: U3 snoRNA
+Macromolecule #2: 5' ETS
+Macromolecule #3: 18S pre-rRNA
+Macromolecule #4: 40S ribosomal protein S1-A
+Macromolecule #5: 40S ribosomal protein S4-A
+Macromolecule #6: 40S ribosomal protein S5
+Macromolecule #7: 40S ribosomal protein S6-A
+Macromolecule #8: 40S ribosomal protein S7-A
+Macromolecule #9: 40S ribosomal protein S8-A
+Macromolecule #10: 40S ribosomal protein S9-A
+Macromolecule #11: 40S ribosomal protein S11-A
+Macromolecule #12: 40S ribosomal protein S12
+Macromolecule #13: 40S ribosomal protein S13
+Macromolecule #14: 40S ribosomal protein S14-A
+Macromolecule #15: 40S ribosomal protein S16-A
+Macromolecule #16: 40S ribosomal protein S18-A
+Macromolecule #17: 40S ribosomal protein S22-B
+Macromolecule #18: 40S ribosomal protein S23-A
+Macromolecule #19: 40S ribosomal protein S24-A
+Macromolecule #20: 40S ribosomal protein S27-A
+Macromolecule #21: 40S ribosomal protein S28-A
+Macromolecule #22: rRNA 2'-O-methyltransferase fibrillarin
+Macromolecule #23: Nucleolar protein 56
+Macromolecule #24: Nucleolar protein 58
+Macromolecule #25: Ribosomal RNA-processing protein 9
+Macromolecule #26: 13 kDa ribonucleoprotein-associated protein
+Macromolecule #27: U3 small nucleolar RNA-associated protein 4
+Macromolecule #28: U3 small nucleolar RNA-associated protein 5
+Macromolecule #29: U3 small nucleolar RNA-associated protein 8
+Macromolecule #30: U3 small nucleolar RNA-associated protein 9
+Macromolecule #31: U3 small nucleolar RNA-associated protein 10
+Macromolecule #32: U3 small nucleolar RNA-associated protein 15
+Macromolecule #33: NET1-associated nuclear protein 1
+Macromolecule #34: Periodic tryptophan protein 2
+Macromolecule #35: U3 small nucleolar RNA-associated protein 12
+Macromolecule #36: U3 small nucleolar RNA-associated protein 13
+Macromolecule #37: U3 small nucleolar RNA-associated protein 18
+Macromolecule #38: U3 small nucleolar RNA-associated protein 21
+Macromolecule #39: U3 small nucleolar RNA-associated protein 6
+Macromolecule #40: Bud site selection protein 21
+Macromolecule #41: U3 small nucleolar RNA-associated protein 7
+Macromolecule #42: U3 small nucleolar RNA-associated protein 11
+Macromolecule #43: U3 small nucleolar RNA-associated protein MPP10
+Macromolecule #44: U3 small nucleolar ribonucleoprotein protein IMP3
+Macromolecule #45: U3 small nucleolar ribonucleoprotein protein IMP4
+Macromolecule #46: Something about silencing protein 10
+Macromolecule #47: Protein SOF1
+Macromolecule #48: rRNA-processing protein FCF2
+Macromolecule #49: rRNA-processing protein FCF1
+Macromolecule #50: Ribosome biogenesis protein ENP2
+Macromolecule #51: U3 small nucleolar ribonucleoprotein protein LCP5
+Macromolecule #52: KRR1 small subunit processome component
+Macromolecule #53: U3 small nucleolar RNA-associated protein 22
+Macromolecule #54: Ribosomal RNA-processing protein 7
+Macromolecule #55: Ribosomal RNA small subunit methyltransferase NEP1
+Macromolecule #56: Ribosome biogenesis protein UTP30
+Macromolecule #57: Ribosome biogenesis protein BMS1
+Macromolecule #58: RNA 3'-terminal phosphate cyclase-like protein
+Macromolecule #59: RNA cytidine acetyltransferase
+Macromolecule #60: Nucleolar complex protein 14
+Macromolecule #61: Nucleolar complex protein 4
+Macromolecule #62: U3 small nucleolar RNA-associated protein 20
+Macromolecule #63: U3 small nucleolar RNA-associated protein 14
+Macromolecule #64: Essential nuclear protein 1
+Macromolecule #65: Pno1
+Macromolecule #66: Protein FAF1
+Macromolecule #67: Regulator of rDNA transcription protein 14
+Macromolecule #68: Protein BFR2
+Macromolecule #69: Unassigned helices
+Macromolecule #70: ZINC ION
+Macromolecule #71: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #72: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 2.0 mg/mL | |||||||||
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Buffer | pH: 7.4 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 10 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. / Pretreatment - Atmosphere: OTHER | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 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: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-32 / Number grids imaged: 10 / Number real images: 18028 / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.5 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |