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- EMDB-21839: SAM-IV riboswitch with SAM -

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Basic information

Entry
Database: EMDB / ID: EMD-21839
TitleSAM-IV riboswitch with SAM
Map data
Sample
  • Complex: SAM-IV riboswitch with SAM
    • RNA: RNA (119-MER)
Keywordsaptamer / riboswitch / RNA
Biological speciesMycobacterium sp. (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.8 Å
AuthorsKappel K / Zhang K
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/Office of the DirectorS10 OD021600 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)R21 AI145647 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)P41GM103832, R01GM079429, U54GM103297, R35 GM112579 United States
CitationJournal: Nat Methods / Year: 2020
Title: Accelerated cryo-EM-guided determination of three-dimensional RNA-only structures.
Authors: Kalli Kappel / Kaiming Zhang / Zhaoming Su / Andrew M Watkins / Wipapat Kladwang / Shanshan Li / Grigore Pintilie / Ved V Topkar / Ramya Rangan / Ivan N Zheludev / Joseph D Yesselman / Wah Chiu / Rhiju Das /
Abstract: The discovery and design of biologically important RNA molecules is outpacing three-dimensional structural characterization. Here, we demonstrate that cryo-electron microscopy can routinely resolve ...The discovery and design of biologically important RNA molecules is outpacing three-dimensional structural characterization. Here, we demonstrate that cryo-electron microscopy can routinely resolve maps of RNA-only systems and that these maps enable subnanometer-resolution coordinate estimation when complemented with multidimensional chemical mapping and Rosetta DRRAFTER computational modeling. This hybrid 'Ribosolve' pipeline detects and falsifies homologies and conformational rearrangements in 11 previously unknown 119- to 338-nucleotide protein-free RNA structures: full-length Tetrahymena ribozyme, hc16 ligase with and without substrate, full-length Vibrio cholerae and Fusobacterium nucleatum glycine riboswitch aptamers with and without glycine, Mycobacterium SAM-IV riboswitch with and without S-adenosylmethionine, and the computer-designed ATP-TTR-3 aptamer with and without AMP. Simulation benchmarks, blind challenges, compensatory mutagenesis, cross-RNA homologies and internal controls demonstrate that Ribosolve can accurately resolve the global architectures of RNA molecules but does not resolve atomic details. These tests offer guidelines for making inferences in future RNA structural studies with similarly accelerated throughput.
History
DepositionApr 20, 2020-
Header (metadata) releaseJul 8, 2020-
Map releaseJul 8, 2020-
UpdateMar 6, 2024-
Current statusMar 6, 2024Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 1.2
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 1.2
  • Imaged by UCSF Chimera
  • Download
  • Surface view with fitted model
  • Atomic models: PDB-6wlr
  • Surface level: 1.2
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_21839.png

Downloads & links

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Map

FileDownload / File: emd_21839.map.gz / Format: CCP4 / Size: 18.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.06 Å/pix.
x 168 pix.
= 178.08 Å		1.06 Å/pix.
x 168 pix.
= 178.08 Å		1.06 Å/pix.
x 168 pix.
= 178.08 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.06 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 1.2 / Movie #1: 1.2
Minimum - Maximum-2.3221495 - 3.6978097
Average (Standard dev.)0.0060318913 (±0.2874408)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions168168168
Spacing168168168
CellA=B=C: 178.07999 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.061.061.06
M x/y/z168168168
origin x/y/z0.0000.0000.000
length x/y/z178.080178.080178.080
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS168168168
D min/max/mean-2.3223.6980.006

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Supplemental data

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Sample components

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Entire : SAM-IV riboswitch with SAM

EntireName: SAM-IV riboswitch with SAM
Components
  • Complex: SAM-IV riboswitch with SAM
    • RNA: RNA (119-MER)

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Supramolecule #1: SAM-IV riboswitch with SAM

SupramoleculeName: SAM-IV riboswitch with SAM / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all / Details: RNA generated by in vitro transcription
Source (natural)Organism: Mycobacterium sp. (bacteria)
Molecular weightTheoretical: 39 KDa

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Macromolecule #1: RNA (119-MER)

MacromoleculeName: RNA (119-MER) / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Mycobacterium sp. (bacteria)
Molecular weightTheoretical: 38.522906 KDa
SequenceString:
GGUCAUGAGU GCCAGCGUCA AGCCCCGGCU UGCUGGCCGG CAACCCUCCA ACCGCGGUGG GGUGCCCCGG GUGAUGACCA GGUUGAGUA GCCGUGACGG CUACGCGGCA AGCGCGGGUC

GENBANK: GENBANK: CP000384.1

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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: 8
GridDetails: unspecified
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 45.6 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

Startup modelType of model: NONE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 225303
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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