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- EMDB-70250: Focused refinement of one subunit or ROOL RNA nanocage -

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Basic information

Entry
Database: EMDB / ID: EMD-70250
TitleFocused refinement of one subunit or ROOL RNA nanocage
Map dataReconstruction of local refinement of one subunit of ROOL RNA nanocage.
Sample
  • Complex: ROOL RNA nanocage - env-120
    • RNA: ROOL RNA
KeywordsROOL / RNA / ncRNA / nanocage
Biological speciessynthetic construct (others)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.1 Å
AuthorsKretsch RC / Wu Y / Das R / Chiu W
Funding support United States, 4 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35 GM122579 United States
Howard Hughes Medical Institute (HHMI) United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)U24 GM129541 United States
National Science Foundation (NSF, United States)2330652 United States
Citation
Journal: Nature / Year: 2025
Title: Naturally ornate RNA-only complexes revealed by cryo-EM.
Authors: Rachael C Kretsch / Yuan Wu / Svetlana A Shabalina / Hyunbin Lee / Grace Nye / Eugene V Koonin / Alex Gao / Wah Chiu / Rhiju Das /
Abstract: The structures of natural RNAs remain poorly characterized and may hold numerous surprises. Here we report three-dimensional structures of three large ornate bacterial RNAs using cryo-electron ...The structures of natural RNAs remain poorly characterized and may hold numerous surprises. Here we report three-dimensional structures of three large ornate bacterial RNAs using cryo-electron microscopy (cryo-EM). GOLLD (Giant, Ornate, Lake- and Lactobacillales-Derived), ROOL (Rumen-Originating, Ornate, Large) and OLE (Ornate Large Extremophilic) RNAs form homo-oligomeric complexes whose stoichiometries are retained at lower concentrations than measured in cells. OLE RNA forms a dimeric complex with long co-axial pipes spanning two monomers. Both GOLLD and ROOL form distinct RNA-only multimeric nanocages with diameters larger than the ribosome, each empty except for a disordered loop. Extensive intramolecular and intermolecular A-minor interactions, kissing loops, an unusual A-A helix and other interactions stabilize the three complexes. Sequence covariation analysis of these large RNAs reveals evolutionary conservation of intermolecular interactions, supporting the biological importance of large, ornate RNA quaternary structures that can assemble without any involvement of proteins.
#1: Journal: BioRxiv / Year: 2025
Title: Naturally ornate RNA-only complexes revealed by cryo-EM
Authors: Kretsch RC / Wu Y / Shabalina SA / Lee H / Nye G / Koonin E / Gao A / Chiu W / Das R
History
DepositionApr 17, 2025-
Header (metadata) releaseApr 30, 2025-
Map releaseApr 30, 2025-
UpdateMay 21, 2025-
Current statusMay 21, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_70250.png

Downloads & links

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Map

FileDownload / File: emd_70250.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationReconstruction of local refinement of one subunit of ROOL RNA nanocage.
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.29 Å/pix.
x 400 pix.
= 516. Å		1.29 Å/pix.
x 400 pix.
= 516. Å		1.29 Å/pix.
x 400 pix.
= 516. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.29 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.25
Minimum - Maximum-0.38009024 - 1.2788725
Average (Standard dev.)0.0019146424 (±0.028626798)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 516.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_70250_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Half map of local refinement of one subunit of ROOL RNA nanocage.

Fileemd_70250_half_map_1.map
AnnotationHalf map of local refinement of one subunit of ROOL RNA nanocage.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Half map of local refinement of one subunit of ROOL RNA nanocage.

Fileemd_70250_half_map_2.map
AnnotationHalf map of local refinement of one subunit of ROOL RNA nanocage.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : ROOL RNA nanocage - env-120

EntireName: ROOL RNA nanocage - env-120
Components
  • Complex: ROOL RNA nanocage - env-120
    • RNA: ROOL RNA

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Supramolecule #1: ROOL RNA nanocage - env-120

SupramoleculeName: ROOL RNA nanocage - env-120 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 1.702 MDa

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Macromolecule #1: ROOL RNA

MacromoleculeName: ROOL RNA / type: rna / ID: 1
Source (natural)Organism: synthetic construct (others)
SequenceString: GGAAUGUUUA UAGACAUAGC CUUGUGUAUG ACUGUCUAAU CAACAGUGCA AGGAAUUAGU UGUGCUCUCA AAAGGAGUUA UGUGAAGGAA AGAUUAAAUG GAUACAUUUA AUUAAGUACA AUUACUACAA AAUCUAUAAU GACUGGGUAA GUCACCCGUA AGAAGGAUGA ...String:
GGAAUGUUUA UAGACAUAGC CUUGUGUAUG ACUGUCUAAU CAACAGUGCA AGGAAUUAGU UGUGCUCUCA AAAGGAGUUA UGUGAAGGAA AGAUUAAAUG GAUACAUUUA AUUAAGUACA AUUACUACAA AAUCUAUAAU GACUGGGUAA GUCACCCGUA AGAAGGAUGA GUUAGUAGAG AUAUAAUAAU AUCUUACAGU UCAACUGAUG UGCAAGUUUA UAAGUAGACG UAGUGUGAAA GACUUAUCGC UAACGCAAUA GACGUUAUCU CGAAAGGAUA AAAGAAACCA AGAAGAUAUA CAAAUUGGUA ACUUGUAUAA GCCCUUGGUA AUACCAAAUA AGUUAGUUCC UCAUGAUGUC GUGGAAACAU GACUAUAAGA UAAGUUGAAU UCGAUAGUAG CCCAAGAGGA AUCAAAUGAG UAAAACAAGU AUUAUAGGUA AAUAUUAUGU UUUUCUAUCU GAAUGAUUGU GCCUUGAAAA AGGUGGGUGA GGUCUGUGGG UAAAAAUUAA UUCCCAUCUG GUUUUGGAUU UUCUUCGGAA AAUUGGUGCA AGAAGCGUGG AGUUGCUAAC CAUUGCUCAG ACUUGUAUCG CCAGUAGCAC GAUAACGAUA UUUAUGAAUG UUGUAAGGUG AAUAAAAGCC UAUGAUUUGU GAACAUUCC

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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

Concentration1.9 mg/mL
BufferpH: 8
Component:
ConcentrationNameFormula
50.0 mMSodium HEPES
10.0 mMMagnesium ChlorideMgCl2
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Number real images: 4462 / Average exposure time: 1.42 sec. / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 105000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 71114
CTF correctionSoftware - Name: cryoSPARC (ver. 4.5.3) / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: INSILICO MODEL
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.5.3) / Software - details: Local refinement / Number images used: 31056
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.5.3) / Software - details: Ab Initio reconstruction multiclass
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.5.3) / Software - details: Homogenous refinement
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementProtocol: AB INITIO MODEL

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