|Entry||Database: EMDB / ID: 8348|
|Title||Cryo-EM Structure of Yeast Pre-40S, Tsr1 region local class "T1"|
|Map data||Pre-40S, local class of Tsr1 region|
|Sample||ribosomal small subunit late-stage assembly intermediate:|
|Source||Saccharomyces cerevisiae (baker's yeast)|
|Method||single particle reconstruction / cryo EM / 10.4 Å resolution|
|Authors||Johnson MC / Ghalei H / Doxtader KA / Karbstein K / Stroupe ME|
|Citation||Journal: Structure / Year: 2017|
Title: Structural Heterogeneity in Pre-40S Ribosomes.
Authors: Matthew C Johnson / Homa Ghalei / Katelyn A Doxtader / Katrin Karbstein / M Elizabeth Stroupe
Abstract: Late-stage 40S ribosome assembly is a highly regulated dynamic process that occurs in the cytoplasm, alongside the full translation machinery. Seven assembly factors (AFs) regulate and facilitate ...Late-stage 40S ribosome assembly is a highly regulated dynamic process that occurs in the cytoplasm, alongside the full translation machinery. Seven assembly factors (AFs) regulate and facilitate maturation, but the mechanisms through which they work remain undetermined. Here, we present a series of structures of the immature small subunit (pre-40S) determined by three-dimensional (3D) cryoelectron microscopy with 3D sorting to assess the molecule's heterogeneity. These structures demonstrate an extensive structural heterogeneity of interface AFs that likely regulates subunit joining during 40S maturation. We also present structural models for the beak and the platform, two regions where the low resolution of previous studies did not allow for localization of AFs and the rRNA, respectively. These models are supported by biochemical analyses using point variants and suggest that maturation of the 18S 3' end is regulated by dissociation of the AF Dim1 from the subunit interface, consistent with previous biochemical analyses.
|Date||Deposition: Aug 26, 2016 / Header (metadata) release: Sep 14, 2016 / Map release: Feb 8, 2017 / Last update: Feb 14, 2018|
|Structure viewer||EM map: |
Downloads & links
|File||emd_8348.map.gz (map file in CCP4 format, 67109 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.62 Å|
CCP4 map header:
-Entire ribosomal small subunit late-stage assembly intermediate
|Entire||Name: ribosomal small subunit late-stage assembly intermediate|
Number of components: 1
-Component #1: protein, ribosomal small subunit late-stage assembly intermediate
|Protein||Name: ribosomal small subunit late-stage assembly intermediate|
Recombinant expression: No
|Source||Species: Saccharomyces cerevisiae (baker's yeast)|
|Specimen||Specimen state: particle / Method: cryo EM|
|Sample solution||pH: 7|
|Vitrification||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: 6 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: DIRECT ELECTRON DE-20 (5k x 3k)|
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