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Yorodumi- EMDB-43027: 60S ribosome biogenesis intermediate (Dbp10 post-catalytic struct... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-43027 | ||||||||||||
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Title | 60S ribosome biogenesis intermediate (Dbp10 post-catalytic structure - Overall map) | ||||||||||||
Map data | 60S ribosome biogenesis intermediate (Dbp10 post-catalytic structure - Overall map) | ||||||||||||
Sample |
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Keywords | Ribosome biogenesis / intermediate DEAD-box ATPase / Ribosome-RNA complex | ||||||||||||
Function / homology | Function and homology information 25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc1p-Noc2p complex / Noc2p-Noc3p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / snoRNA release from pre-rRNA / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity ...25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc1p-Noc2p complex / Noc2p-Noc3p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / snoRNA release from pre-rRNA / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (cytosine-C5-)-methyltransferase activity / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / PeBoW complex / intracellular mRNA localization / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear pre-replicative complex / rRNA base methylation / rRNA primary transcript binding / ATP-dependent activity, acting on RNA / positive regulation of ATP-dependent activity / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA methylation / protein folding chaperone complex / maturation of 5.8S rRNA / hexon binding / pre-mRNA 5'-splice site binding / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome binding / Major pathway of rRNA processing in the nucleolus and cytosol / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / ribosomal large subunit binding / 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 / negative regulation of mRNA splicing, via spliceosome / preribosome, large subunit precursor / nuclear-transcribed mRNA catabolic process / ATPase activator activity / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / DNA replication initiation / 90S preribosome / mRNA transport / regulation of translational fidelity / protein-RNA complex assembly / ribonucleoprotein complex binding / ribosomal subunit export from nucleus / 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) / maturation of LSU-rRNA / translation initiation factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome complex / nuclear periphery / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / cytosolic ribosome assembly / small-subunit processome / macroautophagy / maintenance of translational fidelity / protein catabolic process / rRNA processing / ribosomal small subunit biogenesis / protein transport / ribosome biogenesis / viral capsid / nuclear envelope / ATPase binding / protein-macromolecule adaptor activity / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / RNA helicase activity / negative regulation of translation / rRNA binding / RNA helicase / ribosome / structural constituent of ribosome / translation / cell division / GTPase activity / mRNA binding / chromatin binding / host cell nucleus / GTP binding / nucleolus / ATP hydrolysis activity / mitochondrion / DNA binding / RNA binding / nucleoplasm / ATP binding / identical protein binding Similarity search - Function | ||||||||||||
Biological species | Saccharomyces cerevisiae BY4741 (yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.66 Å | ||||||||||||
Authors | Cruz VE / Weirich CS / Peddada N / Erzberger JP | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: Nat Commun / Year: 2024 Title: The DEAD-box ATPase Dbp10/DDX54 initiates peptidyl transferase center formation during 60S ribosome biogenesis. Authors: Victor E Cruz / Christine S Weirich / Nagesh Peddada / Jan P Erzberger / Abstract: DEAD-box ATPases play crucial roles in guiding rRNA restructuring events during the biogenesis of large (60S) ribosomal subunits, but their precise molecular functions are currently unknown. In this ...DEAD-box ATPases play crucial roles in guiding rRNA restructuring events during the biogenesis of large (60S) ribosomal subunits, but their precise molecular functions are currently unknown. In this study, we present cryo-EM reconstructions of nucleolar pre-60S intermediates that reveal an unexpected, alternate secondary structure within the nascent peptidyl-transferase-center (PTC). Our analysis of three sequential nucleolar pre-60S intermediates reveals that the DEAD-box ATPase Dbp10/DDX54 remodels this alternate base pairing and enables the formation of the rRNA junction that anchors the mature form of the universally conserved PTC A-loop. Post-catalysis, Dbp10 captures rRNA helix H61, initiating the concerted exchange of biogenesis factors during late nucleolar 60S maturation. Our findings show that Dbp10 activity is essential for the formation of the ribosome active site and reveal how this function is integrated with subsequent assembly steps to drive the biogenesis of the large ribosomal subunit. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_43027.map.gz | 166.7 MB | EMDB map data format | |
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Header (meta data) | emd-43027-v30.xml emd-43027.xml | 85.7 KB 85.7 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_43027_fsc.xml | 12.7 KB | Display | FSC data file |
Images | emd_43027.png | 92.3 KB | ||
Filedesc metadata | emd-43027.cif.gz | 20 KB | ||
Others | emd_43027_half_map_1.map.gz emd_43027_half_map_2.map.gz | 140.9 MB 140.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-43027 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-43027 | HTTPS FTP |
-Validation report
Summary document | emd_43027_validation.pdf.gz | 1.1 MB | Display | EMDB validaton report |
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Full document | emd_43027_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | emd_43027_validation.xml.gz | 20.2 KB | Display | |
Data in CIF | emd_43027_validation.cif.gz | 26.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43027 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-43027 | HTTPS FTP |
-Related structure data
Related structure data | 8v87MC 8v83C 8v84C 8v85C C: citing same article (ref.) M: atomic model generated by this map |
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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_43027.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | 60S ribosome biogenesis intermediate (Dbp10 post-catalytic structure - Overall map) | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.08 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: Half Map A
File | emd_43027_half_map_1.map | ||||||||||||
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Annotation | Half Map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half Map B
File | emd_43027_half_map_2.map | ||||||||||||
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Annotation | Half Map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 60S ribosome biogenesis intermediate
+Supramolecule #1: 60S ribosome biogenesis intermediate
+Macromolecule #1: 25S ribosomal RNA
+Macromolecule #2: 5.8S ribosomal RNA
+Macromolecule #4: ITS2 RNA
+Macromolecule #3: ATP-dependent RNA helicase DBP10
+Macromolecule #5: 60S ribosomal subunit assembly/export protein LOC1
+Macromolecule #6: Nucleolar complex protein 2
+Macromolecule #7: Ribosome biogenesis protein BRX1
+Macromolecule #8: 60S ribosomal protein L3
+Macromolecule #9: 60S ribosomal protein L4-A
+Macromolecule #10: ATP-dependent RNA helicase HAS1
+Macromolecule #11: 60S ribosomal protein L6-A
+Macromolecule #12: 60S ribosomal protein L7-A
+Macromolecule #13: Large ribosomal subunit protein eL8A
+Macromolecule #14: Large ribosomal subunit protein uL6A
+Macromolecule #15: Nucleolar complex-associated protein 3
+Macromolecule #16: rRNA-processing protein EBP2
+Macromolecule #17: Proteasome-interacting protein CIC1
+Macromolecule #18: 60S ribosomal protein L13-A
+Macromolecule #19: 60S ribosomal protein L14-A
+Macromolecule #20: Large ribosomal subunit protein eL15A
+Macromolecule #21: Large ribosomal subunit protein uL13A
+Macromolecule #22: 60S ribosomal protein L17-A
+Macromolecule #23: 60S ribosomal protein L18-A
+Macromolecule #24: 60S ribosomal protein L19-A
+Macromolecule #25: Large ribosomal subunit protein eL20A
+Macromolecule #26: 60S ribosomal protein L22-A
+Macromolecule #27: 60S ribosomal protein L23-A
+Macromolecule #28: Ribosome assembly factor MRT4
+Macromolecule #29: 60S ribosomal protein L25
+Macromolecule #30: 60S ribosomal protein L26-A
+Macromolecule #31: 60S ribosomal protein L27-A
+Macromolecule #32: Large ribosomal subunit protein uL1A
+Macromolecule #33: Nucleolar GTP-binding protein 1
+Macromolecule #34: 60S ribosomal protein L30
+Macromolecule #35: 60S ribosomal protein L31-A
+Macromolecule #36: 60S ribosomal protein L32
+Macromolecule #37: 60S ribosomal protein L33-A
+Macromolecule #38: 60S ribosomal protein L34-A
+Macromolecule #39: 60S ribosomal protein L35-A
+Macromolecule #40: 60S ribosomal protein L36-A
+Macromolecule #41: 60S ribosomal protein L37-A
+Macromolecule #42: 60S ribosomal protein L38
+Macromolecule #43: 60S ribosome subunit biogenesis protein NIP7
+Macromolecule #44: Ribosome biogenesis protein ERB1
+Macromolecule #45: Pescadillo homolog
+Macromolecule #46: Ribosome biogenesis protein 15
+Macromolecule #47: YTM1 isoform 1
+Macromolecule #48: 25S rRNA (cytosine(2870)-C(5))-methyltransferase
+Macromolecule #49: Ribosome biogenesis protein NSA2
+Macromolecule #50: Nuclear GTP-binding protein NUG1
+Macromolecule #51: Ribosome biogenesis protein RLP7
+Macromolecule #52: Ribosome biogenesis protein RLP24
+Macromolecule #53: Nucleolar protein 16
+Macromolecule #54: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase
+Macromolecule #55: Eukaryotic translation initiation factor 6
+Macromolecule #56: UPF0642 protein YBL028C
+Macromolecule #57: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #58: MAGNESIUM ION
+Macromolecule #59: GUANOSINE-5'-DIPHOSPHATE
+Macromolecule #60: POTASSIUM ION
+Macromolecule #61: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.5 mg/mL | |||||||||||||||
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Buffer | pH: 8 Component:
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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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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 39.3 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.9 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |