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- EMDB-44457: 80S ribosome with angiogenin and ternary complex in rabbit reticu... -

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Basic information

Entry
Database: EMDB / ID: EMD-44457
Title80S ribosome with angiogenin and ternary complex in rabbit reticulocyte lysates
Map data80S ribosome with angiogenin and ternary complex in RRL, masked map used for figures
Sample
  • Complex: 80S ribosome with angiogenin and ternary complex in rabbit reticulocyte lysate
    • Complex: 80S ribosome
    • Complex: angiogenin
KeywordsAngiogenin / RNase / RIBOSOME
Biological speciesOryctolagus cuniculus (rabbit) / Homo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.36 Å
AuthorsLoveland AB / Korostelev AA
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01 GM127094 United States
CitationJournal: Nature / Year: 2024
Title: Structural mechanism of angiogenin activation by the ribosome.
Authors: Anna B Loveland / Cha San Koh / Robin Ganesan / Allan Jacobson / Andrei A Korostelev /
Abstract: Angiogenin, an RNase-A-family protein, promotes angiogenesis and has been implicated in cancer, neurodegenerative diseases and epigenetic inheritance. After activation during cellular stress, ...Angiogenin, an RNase-A-family protein, promotes angiogenesis and has been implicated in cancer, neurodegenerative diseases and epigenetic inheritance. After activation during cellular stress, angiogenin cleaves tRNAs at the anticodon loop, resulting in translation repression. However, the catalytic activity of isolated angiogenin is very low, and the mechanisms of the enzyme activation and tRNA specificity have remained a puzzle. Here we identify these mechanisms using biochemical assays and cryogenic electron microscopy (cryo-EM). Our study reveals that the cytosolic ribosome is the activator of angiogenin. A cryo-EM structure features angiogenin bound in the A site of the 80S ribosome. The C-terminal tail of angiogenin is rearranged by interactions with the ribosome to activate the RNase catalytic centre, making the enzyme several orders of magnitude more efficient in tRNA cleavage. Additional 80S-angiogenin structures capture how tRNA substrate is directed by the ribosome into angiogenin's active site, demonstrating that the ribosome acts as the specificity factor. Our findings therefore suggest that angiogenin is activated by ribosomes with a vacant A site, the abundance of which increases during cellular stress. These results may facilitate the development of therapeutics to treat cancer and neurodegenerative diseases.
History
DepositionApr 12, 2024-
Header (metadata) releaseJul 24, 2024-
Map releaseJul 24, 2024-
UpdateJul 24, 2024-
Current statusJul 24, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_44457.png

Downloads & links

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Map

FileDownload / File: emd_44457.map.gz / Format: CCP4 / Size: 857.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation80S ribosome with angiogenin and ternary complex in RRL, masked map used for figures
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.83 Å/pix.
x 608 pix.
= 504.64 Å		0.83 Å/pix.
x 608 pix.
= 504.64 Å		0.83 Å/pix.
x 608 pix.
= 504.64 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.83 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 4.0
Minimum - Maximum-10.168813999999999 - 23.220103999999999
Average (Standard dev.)0.11252468 (±0.8446425)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions608608608
Spacing608608608
CellA=B=C: 504.63998 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_44457_msk_1.map
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AxesZYX

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Additional map: 80S ribosome with angiogenin and ternary complex in RRL, original map

Fileemd_44457_additional_1.map
Annotation80S ribosome with angiogenin and ternary complex in RRL, original map
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Half map: 80S ribosome with angiogenin and ternary complex in RRL, half map 1

Fileemd_44457_half_map_1.map
Annotation80S ribosome with angiogenin and ternary complex in RRL, half map 1
Projections & Slices
AxesZYX

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Histogram

Histogram (log scale)

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Half map: 80S ribosome with angiogenin and ternary complex in RRL, half map 2

Fileemd_44457_half_map_2.map
Annotation80S ribosome with angiogenin and ternary complex in RRL, half map 2
Projections & Slices
AxesZYX

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Histogram

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Sample components

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Entire : 80S ribosome with angiogenin and ternary complex in rabbit reticu...

EntireName: 80S ribosome with angiogenin and ternary complex in rabbit reticulocyte lysate
Components
  • Complex: 80S ribosome with angiogenin and ternary complex in rabbit reticulocyte lysate
    • Complex: 80S ribosome
    • Complex: angiogenin

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Supramolecule #1: 80S ribosome with angiogenin and ternary complex in rabbit reticu...

SupramoleculeName: 80S ribosome with angiogenin and ternary complex in rabbit reticulocyte lysate
type: complex / ID: 1 / Parent: 0
Molecular weightTheoretical: 4.5 MDa

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Supramolecule #2: 80S ribosome

SupramoleculeName: 80S ribosome / type: complex / ID: 2 / Parent: 1
Source (natural)Organism: Oryctolagus cuniculus (rabbit) / Tissue: Reticulocyte

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Supramolecule #3: angiogenin

SupramoleculeName: angiogenin / type: complex / ID: 3 / Parent: 1
Source (natural)Organism: Homo sapiens (human)

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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
GridModel: Quantifoil R2/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV
Details33% rabbit reticulocyte lysate

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 30.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.1 µm / Nominal defocus min: 0.3 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 6216
Startup modelType of model: EMDB MAP
EMDB ID:

4729

Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.36 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 1960
Initial angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: cisTEM
Final angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: cisTEM
Final 3D classificationNumber classes: 6 / Avg.num./class: 1000 / Software - Name: FREALIGN
FSC plot (resolution estimation)

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