|Entry||Database: EMDB / ID: 0330|
|Title||Rea1 Wild type ADP state (AAA+ ring + stem)|
|Sample||Rea1 (MIDASIN) ring with ADP:|
|Source||Saccharomyces cerevisiae (baker's yeast)|
|Method||single particle reconstruction / cryo EM / 4.4 Å resolution|
|Authors||Sosnowski P / Urnavicius L / Boland A / Fagiewicz R / Busselez J / Papai G / Schmidt H|
|Citation||Journal: Elife / Year: 2018|
Title: The CryoEM structure of the ribosome maturation factor Rea1.
Authors: Piotr Sosnowski / Linas Urnavicius / Andreas Boland / Robert Fagiewicz / Johan Busselez / Gabor Papai / Helgo Schmidt
Abstract: The biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly ...The biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly factors. The ~5000 amino-acid AAA+ ATPase Rea1 (or Midasin) generates force to mechanically remove assembly factors from pre-60S particles, which promotes their export to the cytosol. Here we present three Rea1 cryoEM structures. We visualise the Rea1 engine, a hexameric ring of AAA+ domains, and identify an α-helical bundle of AAA2 as a major ATPase activity regulator. The α-helical bundle interferes with nucleotide-induced conformational changes that create a docking site for the substrate binding MIDAS domain on the AAA +ring. Furthermore, we reveal the architecture of the Rea1 linker, which is involved in force generation and extends from the AAA+ ring. The data presented here provide insights into the mechanism of one of the most complex ribosome maturation factors.
|Date||Deposition: Oct 31, 2018 / Header (metadata) release: Dec 5, 2018 / Map release: Dec 5, 2018 / Last update: Dec 5, 2018|
|Structure viewer||EM map: |
Downloads & links
|File||emd_0330.map.gz (map file in CCP4 format, 226493 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.1 Å|
CCP4 map header:
-Entire Rea1 (MIDASIN) ring with ADP
|Entire||Name: Rea1 (MIDASIN) ring with ADP / Number of components: 1|
-Component #1: protein, Rea1 (MIDASIN) ring with ADP
|Protein||Name: Rea1 (MIDASIN) ring with ADP / Recombinant expression: No|
|Source||Species: Saccharomyces cerevisiae (baker's yeast)|
|Source (engineered)||Expression System: Saccharomyces cerevisiae (baker's yeast)|
|Specimen||Specimen state: particle / Method: cryo EM|
|Sample solution||Specimen conc.: 1 mg/ml / Buffer solution: ADP was added 5 minute before the plunging / pH: 7.2|
|Vitrification||Instrument: HOMEMADE PLUNGER / 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: 46.2 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 105000.0 X (nominal), 105000.0 X (calibrated)|
Cs: 0.01 mm / Imaging mode: BRIGHT FIELD / Defocus: 1800.0 - 3400.0 nm / Energy filter: GIF Quantum LS
|Specimen Holder||Model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Image acquisition||Number of digital images: 23230 / Sampling size: 5 microns|
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