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- EMDB-8350: Cryo-EM Structure of Yeast Pre-40S, Rio2 Region Local Class "R1" -

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Basic information

Entry
Database: EMDB / ID: EMD-8350
TitleCryo-EM Structure of Yeast Pre-40S, Rio2 Region Local Class "R1"
Map dataPre-40S, Rio2 region local class "R1"
Sample
  • Complex: ribosomal small subunit late-stage assembly intermediate
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 10.1 Å
AuthorsJohnson MC / Ghalei H / Doxtader KA / Karbstein K / Stroupe ME
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM117093 United States
National Science Foundation (NSF, United States)MCB1149763 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM086451 United States
CitationJournal: 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.
History
DepositionAug 26, 2016-
Header (metadata) releaseSep 14, 2016-
Map releaseFeb 8, 2017-
UpdateDec 25, 2019-
Current statusDec 25, 2019Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.3
  • Imaged by UCSF Chimera
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  • Surface view colored by height
  • Surface level: 0.3
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_8350.png

Downloads & links

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Map

FileDownload / File: emd_8350.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationPre-40S, Rio2 region local class "R1"
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.62 Å/pix.
x 256 pix.
= 414.72 Å		1.62 Å/pix.
x 256 pix.
= 414.72 Å		1.62 Å/pix.
x 256 pix.
= 414.72 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.62 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.3 / Movie #1: 0.3
Minimum - Maximum-0.090097174 - 0.6950206
Average (Standard dev.)0.0037951998 (±0.034281217)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 414.72 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.621.621.62
M x/y/z256256256
origin x/y/z0.0000.0000.000
length x/y/z414.720414.720414.720
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ281156
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS256256256
D min/max/mean-0.0900.6950.004

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Supplemental data

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Sample components

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Entire : ribosomal small subunit late-stage assembly intermediate

EntireName: ribosomal small subunit late-stage assembly intermediate
Components
  • Complex: ribosomal small subunit late-stage assembly intermediate

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Supramolecule #1: ribosomal small subunit late-stage assembly intermediate

SupramoleculeName: ribosomal small subunit late-stage assembly intermediate
type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: DIRECT ELECTRON DE-20 (5k x 3k) / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 16485
Startup modelType of model: EMDB MAP
EMDB ID:

1927

Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 10.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 131842
Initial angle assignmentType: RANDOM ASSIGNMENT
Final angle assignmentType: ANGULAR RECONSTITUTION
FSC plot (resolution estimation)

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