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Open data
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Basic information
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Title | Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement | |||||||||
![]() | Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement - local resolution filtered | |||||||||
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![]() | Human / ribosome biogenesis / NLE1 / RIX1 complex / 5S RNP / RIBOSOME | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.39 Å | |||||||||
![]() | Thoms M / Denk T / Beckmann R | |||||||||
Funding support | European Union, 1 items
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![]() | ![]() Title: Highly conserved ribosome biogenesis pathways between human and yeast revealed by the MDN1-NLE1 interaction and NLE1 containing pre-60S subunits. Authors: Federica Fiorentino / Matthias Thoms / Klemens Wild / Timo Denk / Jingdong Cheng / Jakub Zeman / Irmgard Sinning / Ed Hurt / Roland Beckmann / ![]() ![]() Abstract: The assembly of ribosomal subunits, primarily occurring in the nucleolar and nuclear compartments, is a highly complex process crucial for cellular function. This study reveals the conservation of ...The assembly of ribosomal subunits, primarily occurring in the nucleolar and nuclear compartments, is a highly complex process crucial for cellular function. This study reveals the conservation of ribosome biogenesis between yeast and humans, illustrated by the structural similarities of ribosomal subunit intermediates. By using X-ray crystallography and cryo-EM, the interaction between the human AAA+ ATPase MDN1 and the 60S assembly factor NLE1 is compared with the yeast homologs Rea1 and Rsa4. The MDN1-MIDAS and NLE1-Ubl complex structure at 2.3 Å resolution mirrors the highly conserved interaction patterns observed in yeast. Moreover, human pre-60S intermediates bound to the dominant negative NLE1-E85A mutant revealed at 2.8 Å resolution an architecture that largely matched the equivalent yeast structures. Conformation of rRNA, assembly factors and their interaction networks are highly conserved. Additionally, novel human pre-60S intermediates with a non-rotated 5S RNP and processed ITS2/foot structure but incomplete intersubunit surface were identified to be similar to counterparts observed in yeast. These findings confirm that the MDN1-NLE1-driven transition phase of the 60S assembly is essentially identical, supporting the idea that ribosome biogenesis is a highly conserved process across eukaryotic cells, employing an evolutionary preservation of ribosomal assembly mechanisms. #1: ![]() Title: Real-space refinement in PHENIX for cryo-EM and crystallography Authors: Adams PD | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 6.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 20.5 KB 20.5 KB | Display Display | ![]() |
Images | ![]() | 74.9 KB | ||
Filedesc metadata | ![]() | 4.1 KB | ||
Others | ![]() ![]() ![]() | 237.4 MB 443 MB 443 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 919.2 KB | Display | ![]() |
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Full document | ![]() | 918.8 KB | Display | |
Data in XML | ![]() | 18.5 KB | Display | |
Data in CIF | ![]() | 22.2 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||
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Annotation | Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement - local resolution filtered | ||||||||||||||||||||||||||||||||||||
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
-Additional map: Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement
File | emd_19309_additional_1.map | ||||||||||||
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Annotation | Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Human pre-60S - State 3 - 5S RNP...
File | emd_19309_half_map_1.map | ||||||||||||
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Annotation | Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement - half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Human pre-60S - State 3 - 5S RNP...
File | emd_19309_half_map_2.map | ||||||||||||
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Annotation | Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement - half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement
Entire | Name: Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement |
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Components |
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-Supramolecule #1: Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement
Supramolecule | Name: Human pre-60S - State 3 - 5S RNP - NLE1 - local refinement type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#40 |
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Source (natural) | Organism: ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
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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) / Average electron dose: 43.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: NONE |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.39 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 33891 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |