+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-14992 | ||||||||||||
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Title | FAP-80S Complex - Non-rotated state with empty A site | ||||||||||||
Map data | FAP-80S - Non-rotated state - Empty A site | ||||||||||||
Sample |
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Biological species | Saccharomyces cerevisiae (brewer's yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | ||||||||||||
Authors | Ikeuchi K / Buschauer R / Berninghausen O / Becker T / Beckmann R | ||||||||||||
Funding support | Germany, European Union, 3 items
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Citation | Journal: Mol Cell / Year: 2022 Title: Sensing of individual stalled 80S ribosomes by Fap1 for nonfunctional rRNA turnover. Authors: Sihan Li / Ken Ikeuchi / Misaki Kato / Robert Buschauer / Takato Sugiyama / Shungo Adachi / Hideo Kusano / Tohru Natsume / Otto Berninghausen / Yoshitaka Matsuo / Thomas Becker / Roland ...Authors: Sihan Li / Ken Ikeuchi / Misaki Kato / Robert Buschauer / Takato Sugiyama / Shungo Adachi / Hideo Kusano / Tohru Natsume / Otto Berninghausen / Yoshitaka Matsuo / Thomas Becker / Roland Beckmann / Toshifumi Inada / Abstract: Cells can respond to stalled ribosomes by sensing ribosome collisions and employing quality control pathways. How ribosome stalling is resolved without collisions, however, has remained elusive. ...Cells can respond to stalled ribosomes by sensing ribosome collisions and employing quality control pathways. How ribosome stalling is resolved without collisions, however, has remained elusive. Here, focusing on noncolliding stalling exhibited by decoding-defective ribosomes, we identified Fap1 as a stalling sensor triggering 18S nonfunctional rRNA decay via polyubiquitination of uS3. Ribosome profiling revealed an enrichment of Fap1 at the translation initiation site but also an association with elongating individual ribosomes. Cryo-EM structures of Fap1-bound ribosomes elucidated Fap1 probing the mRNA simultaneously at both the entry and exit channels suggesting an mRNA stasis sensing activity, and Fap1 sterically hinders the formation of canonical collided di-ribosomes. Our findings indicate that individual stalled ribosomes are the potential signal for ribosome dysfunction, leading to accelerated turnover of the ribosome itself. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_14992.map.gz | 241.2 MB | EMDB map data format | |
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Header (meta data) | emd-14992-v30.xml emd-14992.xml | 24.4 KB 24.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_14992_fsc.xml | 16.9 KB | Display | FSC data file |
Images | emd_14992.png | 165.1 KB | ||
Others | emd_14992_half_map_1.map.gz emd_14992_half_map_2.map.gz | 337.9 MB 338 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-14992 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-14992 | HTTPS FTP |
-Validation report
Summary document | emd_14992_validation.pdf.gz | 802.3 KB | Display | EMDB validaton report |
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Full document | emd_14992_full_validation.pdf.gz | 801.9 KB | Display | |
Data in XML | emd_14992_validation.xml.gz | 23.7 KB | Display | |
Data in CIF | emd_14992_validation.cif.gz | 31.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-14992 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-14992 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_14992.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | FAP-80S - Non-rotated state - Empty A site | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.045 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: Half map 1
File | emd_14992_half_map_1.map | ||||||||||||
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Annotation | Half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map 2
File | emd_14992_half_map_2.map | ||||||||||||
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Annotation | Half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Fap1-bound yeast 80S ribosome in non-rotated state with empty A site
Entire | Name: Fap1-bound yeast 80S ribosome in non-rotated state with empty A site |
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Components |
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-Supramolecule #1: Fap1-bound yeast 80S ribosome in non-rotated state with empty A site
Supramolecule | Name: Fap1-bound yeast 80S ribosome in non-rotated state with empty A site type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1-#85 |
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Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: W303 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 43.4 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |