|Entry||Database: EMDB / ID: 3892|
|Title||State F (Rix1-TAP Rpf2-Flag) - Visualizing the assembly pathway of nucleolar pre-60S ribosomes|
|Map data||State F (Rix1-TAP Rpf2-Flag) - Visualizing the assembly pathway of nucleolar pre-60S ribosmes|
|Sample||Large subunit biogenesis particle purified via Rix1-TAP and Rpf2-Flag:|
|Source||Saccharomyces cerevisiae (baker's yeast)|
|Method||single particle reconstruction / cryo EM / 3.3 Å resolution|
|Authors||Kater L / Thoms M / Barrio-Garcia C / Cheng J / Ismail S / Ahmed YL / Bange G / Kressler D / Berninghausen O / Sinning I / Hurt E / Beckmann R|
|Citation||Journal: Cell / Year: 2017|
Title: Visualizing the Assembly Pathway of Nucleolar Pre-60S Ribosomes.
Authors: Lukas Kater / Matthias Thoms / Clara Barrio-Garcia / Jingdong Cheng / Sherif Ismail / Yasar Luqman Ahmed / Gert Bange / Dieter Kressler / Otto Berninghausen / Irmgard Sinning / Ed Hurt / Roland Beckmann
Abstract: Eukaryotic 60S ribosomal subunits are comprised of three rRNAs and ∼50 ribosomal proteins. The initial steps of their formation take place in the nucleolus, but, owing to a lack of structural ...Eukaryotic 60S ribosomal subunits are comprised of three rRNAs and ∼50 ribosomal proteins. The initial steps of their formation take place in the nucleolus, but, owing to a lack of structural information, this process is poorly understood. Using cryo-EM, we solved structures of early 60S biogenesis intermediates at 3.3 Å to 4.5 Å resolution, thereby providing insights into their sequential folding and assembly pathway. Besides revealing distinct immature rRNA conformations, we map 25 assembly factors in six different assembly states. Notably, the Nsa1-Rrp1-Rpf1-Mak16 module stabilizes the solvent side of the 60S subunit, and the Erb1-Ytm1-Nop7 complex organizes and connects through Erb1's meandering N-terminal extension, eight assembly factors, three ribosomal proteins, and three 25S rRNA domains. Our structural snapshots reveal the order of integration and compaction of the six major 60S domains within early nucleolar 60S particles developing stepwise from the solvent side around the exit tunnel to the central protuberance.
|Date||Deposition: Sep 29, 2017 / Header (metadata) release: Dec 20, 2017 / Map release: Dec 27, 2017 / Last update: Dec 27, 2017|
|Structure viewer||EM map: |
Downloads & links
|File||emd_3892.map.gz (map file in CCP4 format, 296353 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.084 Å|
CCP4 map header:
-Entire Large subunit biogenesis particle purified via Rix1-TAP and Rpf2-Flag
|Entire||Name: Large subunit biogenesis particle purified via Rix1-TAP and Rpf2-Flag|
Number of components: 1
-Component #1: protein, Large subunit biogenesis particle purified via Rix1-TAP ...
|Protein||Name: Large subunit biogenesis particle purified via Rix1-TAP and Rpf2-Flag|
Recombinant expression: No
|Source||Species: Saccharomyces cerevisiae (baker's yeast)|
|Specimen||Specimen state: particle / Method: cryo EM|
|Sample solution||pH: 7.5|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 27 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Cs: 2.7 mm / Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: FEI FALCON II (4k x 4k)|
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