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Yorodumi- EMDB-24271: State E2 nucleolar 60S ribosomal biogenesis intermediate - Spb1-M... -
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Open data
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Basic information
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| Title | State E2 nucleolar 60S ribosomal biogenesis intermediate - Spb1-MTD locally refined map | ||||||||||||
Map data | State E2 Spb1-MTD locally refined map | ||||||||||||
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Keywords | ribosome biogenesis / DEAD-box ATPases / methyltransferase / nucleolus / RIBOSOME | ||||||||||||
| Function / homology | Function and homology information25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc1p-Noc2p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / Noc2p-Noc3p complex / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / rRNA (cytosine-C5-)-methyltransferase activity / positive regulation of ATP-dependent activity / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc1p-Noc2p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / Noc2p-Noc3p complex / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / rRNA (cytosine-C5-)-methyltransferase activity / positive regulation of ATP-dependent activity / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA base methylation / rRNA methylation / maturation of 5.8S rRNA / Major pathway of rRNA processing in the nucleolus and cytosol / ribosomal large subunit binding / 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) / preribosome, large subunit precursor / ATPase activator activity / Formation of a pool of free 40S subunits / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / regulation of translational fidelity / ribosomal subunit export from nucleus / maturation of LSU-rRNA / translation initiation factor activity / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / cytosolic ribosome assembly / maintenance of translational fidelity / rRNA processing / protein transport / ATPase binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / structural constituent of ribosome / translation / GTPase activity / mRNA binding / GTP binding / nucleolus / mitochondrion / RNA binding / nucleoplasm / nucleus / cytoplasm / cytosol Similarity search - Function | ||||||||||||
| Biological species | ![]() | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.13 Å | ||||||||||||
Authors | Cruz VE / Sekulski K / Peddada N / Erzberger JP | ||||||||||||
| Funding support | United States, 3 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2022Title: Sequence-specific remodeling of a topologically complex RNP substrate by Spb4. Authors: Victor Emmanuel Cruz / Kamil Sekulski / Nagesh Peddada / Carolin Sailer / Sahana Balasubramanian / Christine S Weirich / Florian Stengel / Jan P Erzberger / ![]() Abstract: DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular ...DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular roles, most notably in driving large-scale RNA remodeling steps during the assembly of ribonucleoproteins (RNPs). We describe cryo-EM structures of 60S ribosomal biogenesis intermediates that reveal how context-specific RNA unwinding by the DEAD-box ATPase Spb4 results in extensive, sequence-specific remodeling of rRNA secondary structure. Multiple cis and trans interactions stabilize Spb4 in a post-catalytic, high-energy intermediate that drives the organization of the three-way junction at the base of rRNA domain IV. This mechanism explains how limited strand separation by DEAD-box ATPases is leveraged to provide non-equilibrium directionality and ensure efficient and accurate RNP assembly. | ||||||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_24271.map.gz | 254.4 MB | EMDB map data format | |
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| Header (meta data) | emd-24271-v30.xml emd-24271.xml | 34.5 KB 34.5 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_24271_fsc.xml | 14.9 KB | Display | FSC data file |
| Images | emd_24271.png | 84.9 KB | ||
| Filedesc metadata | emd-24271.cif.gz | 8.2 KB | ||
| Others | emd_24271_half_map_1.map.gz emd_24271_half_map_2.map.gz | 242.6 MB 242.9 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-24271 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-24271 | HTTPS FTP |
-Validation report
| Summary document | emd_24271_validation.pdf.gz | 877.3 KB | Display | EMDB validaton report |
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| Full document | emd_24271_full_validation.pdf.gz | 876.9 KB | Display | |
| Data in XML | emd_24271_validation.xml.gz | 22.6 KB | Display | |
| Data in CIF | emd_24271_validation.cif.gz | 29.6 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-24271 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-24271 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7nafMC ![]() 7nacC ![]() 7nadC ![]() 7r6kC ![]() 7r6qC ![]() 7r72C ![]() 7r7aC ![]() 7r7cC ![]() 7u0hC C: citing same article ( M: atomic model generated by this map |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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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_24271.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | State E2 Spb1-MTD locally refined 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: State E2 Spb1-MTD locally refined half map 2
| File | emd_24271_half_map_1.map | ||||||||||||
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| Annotation | State E2 Spb1-MTD locally refined half map 2 | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: State E2 Spb1-MTD locally refined half map 1
| File | emd_24271_half_map_2.map | ||||||||||||
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| Annotation | State E2 Spb1-MTD locally refined half map 1 | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4.
+Supramolecule #1: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4.
+Macromolecule #1: 25S rRNA
+Macromolecule #2: Ribosomal RNA-processing protein 17
+Macromolecule #3: Nucleolar complex protein 2
+Macromolecule #4: 60S ribosomal protein L23-A
+Macromolecule #5: Ribosome biogenesis protein RLP24
+Macromolecule #6: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase
+Macromolecule #7: 60S ribosome biogenesis factor Nog1
+Macromolecule #8: 60S ribosomal protein L31-A
+Macromolecule #9: 60S ribosomal protein L22-A
+Macromolecule #10: 60S ribosome ribosomal protein L19A
+Macromolecule #11: 60S ribosomal protein L30
+Macromolecule #12: 60S ribosomal protein L3
+Macromolecule #13: UPF0642 protein YBL028C
+Macromolecule #14: Nuclear GTP-binding protein NUG1
+Macromolecule #15: Eukaryotic translation initiation factor 6
+Macromolecule #16: 25S rRNA (cytosine(2870)-C(5))-methyltransferase
+Macromolecule #17: S-ADENOSYL-L-HOMOCYSTEINE
+Macromolecule #18: water
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.45 mg/mL | ||||||||||||||||||
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| Buffer | pH: 8 Component:
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| Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 0.3 | ||||||||||||||||||
| 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 K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 4523 / Average exposure time: 0.05 sec. / Average electron dose: 1.2 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.9 µm / 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 |
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Image processing
-Atomic model buiding 1
| Refinement | Protocol: RIGID BODY FIT |
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| Output model | ![]() PDB-7naf: |
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About Yorodumi



Keywords
Authors
United States, 3 items
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Z (Sec.)
Y (Row.)
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FIELD EMISSION GUN


