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- EMDB-36779: The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 ... -

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Basic information

Entry
Database: EMDB / ID: EMD-36779
TitleThe Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 with 10-nt TRS at symmetric apo state
Map data
Sample
  • Complex: The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 with 10-nt TRS at symmetric apo state
    • RNA: RNA (543-MER)
  • Ligand: MAGNESIUM ION
KeywordsORF-less Group IIC intron / ribozymes / RNA
Biological speciesAnoxybacillus pushchinoensis (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.41 Å
AuthorsZhu HZ / Liu JJG
Funding support China, 1 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC) China
CitationJournal: Science / Year: 2024
Title: Hydrolytic endonucleolytic ribozyme (HYER) is programmable for sequence-specific DNA cleavage.
Authors: Zi-Xian Liu / Shouyue Zhang / Han-Zhou Zhu / Zhi-Hang Chen / Yun Yang / Long-Qi Li / Yuan Lei / Yun Liu / Dan-Yuan Li / Ao Sun / Cheng-Ping Li / Shun-Qing Tan / Gao-Li Wang / Jie-Yi Shen / ...Authors: Zi-Xian Liu / Shouyue Zhang / Han-Zhou Zhu / Zhi-Hang Chen / Yun Yang / Long-Qi Li / Yuan Lei / Yun Liu / Dan-Yuan Li / Ao Sun / Cheng-Ping Li / Shun-Qing Tan / Gao-Li Wang / Jie-Yi Shen / Shuai Jin / Caixia Gao / Jun-Jie Gogo Liu /
Abstract: Ribozymes are catalytic RNAs with diverse functions including self-splicing and polymerization. This work aims to discover natural ribozymes that behave as hydrolytic and sequence-specific DNA ...Ribozymes are catalytic RNAs with diverse functions including self-splicing and polymerization. This work aims to discover natural ribozymes that behave as hydrolytic and sequence-specific DNA endonucleases, which could be repurposed as DNA manipulation tools. Focused on bacterial group II-C introns, we found that many systems without intron-encoded protein propagate multiple copies in their resident genomes. These introns, named HYdrolytic Endonucleolytic Ribozymes (HYERs), cleaved RNA, single-stranded DNA, bubbled double-stranded DNA (dsDNA), and plasmids in vitro. HYER1 generated dsDNA breaks in the mammalian genome. Cryo-electron microscopy analysis revealed a homodimer structure for HYER1, where each monomer contains a Mg-dependent hydrolysis pocket and captures DNA complementary to the target recognition site (TRS). Rational designs including TRS extension, recruiting sequence insertion, and heterodimerization yielded engineered HYERs showing improved specificity and flexibility for DNA manipulation.
History
DepositionJul 10, 2023-
Header (metadata) releaseMay 1, 2024-
Map releaseMay 1, 2024-
UpdateMay 15, 2024-
Current statusMay 15, 2024Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_36779.png

Downloads & links

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Map

FileDownload / File: emd_36779.map.gz / Format: CCP4 / Size: 476.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.0825 Å
Density
Contour LevelBy AUTHOR: 0.5
Minimum - Maximum-0.76134616 - 1.7406143
Average (Standard dev.)-0.00039204452 (±0.042870376)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions500500500
Spacing500500500
CellA=B=C: 541.25 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_36779_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_36779_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 ...

EntireName: The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 with 10-nt TRS at symmetric apo state
Components
  • Complex: The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 with 10-nt TRS at symmetric apo state
    • RNA: RNA (543-MER)
  • Ligand: MAGNESIUM ION

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Supramolecule #1: The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 ...

SupramoleculeName: The Anoxybacillus pushchinoensis ORF-less Group IIC Intron HYER1 with 10-nt TRS at symmetric apo state
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1
Source (natural)Organism: Anoxybacillus pushchinoensis (bacteria)

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Macromolecule #1: RNA (543-MER)

MacromoleculeName: RNA (543-MER) / type: rna / ID: 1 / Number of copies: 2
Source (natural)Organism: Anoxybacillus pushchinoensis (bacteria)
Molecular weightTheoretical: 206.396656 KDa
SequenceString: GUGCGCUCGG CAUGGGUGCA AUCUCUAGGG UGAAAGUCCC GAACUGCGAA GGCAGAAGUA GCAGUUAGCU UAACGCAAGG GUGUCCGUG GUGACGCGGA AUCUGAAGGA AGCGGGCGGC AAACUUCCGG UCUGAGGAAC ACGAACUUCA UAUAAGGCUA G GUAUCAUU ...String:
GUGCGCUCGG CAUGGGUGCA AUCUCUAGGG UGAAAGUCCC GAACUGCGAA GGCAGAAGUA GCAGUUAGCU UAACGCAAGG GUGUCCGUG GUGACGCGGA AUCUGAAGGA AGCGGGCGGC AAACUUCCGG UCUGAGGAAC ACGAACUUCA UAUAAGGCUA G GUAUCAUU GGAUGAGUUU GCCGCUAAGA CAAAACAAAG UCCUUUCUGC CGAAGGUGAU ACAGAGUAAA UGAAGCAGAU AG AUGGAAG GAAAGAUUGU ACUCUUACCC GAGGAGGUCU GAUGGAUACG UGAAGUGCGC UUCAUAACCU ACUUAGUGAU AAG UAACUG AACCAUCAGA AGUCAGCAGA GGUCAUAGUA CGAAUCGGUC UAGAACGAUU CGGAAGGACU GAACAAUCAA GAGA AAAUA GCCCUUGGCA UUCAGUACGU CAUGAUGAAC ACAGAAAACA UGGUACCUCC CAAGAGAAAG GAAACGGUGA AUCCC GUGG GAAUCUUUUG GAGGGUGGAG UGACGACUGG CAUAAGAAGA UCAGCUAUUU ACGGAAGGAA GCUUGCGUCA UUAUCU UGA UUGAACCGCC GUAUACGGAA CCGUACGUAC GGUGGUGUGA GAGGACGGAG GUUAAUCACC UCCUCCUACU CGAU

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Macromolecule #2: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 2 / Number of copies: 2 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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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: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.5 µm
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: OTHER
Initial angle assignmentType: OTHER
Final angle assignmentType: OTHER
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.41 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 166165

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