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TitleSub-3-Å cryo-EM structure of RNA enabled by engineered homomeric self-assembly.
Journal, issue, pagesNat Methods, Vol. 19, Issue 5, Page 576-585, Year 2022
Publish dateMay 2, 2022
AuthorsDi Liu / François A Thélot / Joseph A Piccirilli / Maofu Liao / Peng Yin /
PubMed AbstractHigh-resolution structural studies are essential for understanding the folding and function of diverse RNAs. Herein, we present a nanoarchitectural engineering strategy for efficient structural ...High-resolution structural studies are essential for understanding the folding and function of diverse RNAs. Herein, we present a nanoarchitectural engineering strategy for efficient structural determination of RNA-only structures using single-particle cryogenic electron microscopy (cryo-EM). This strategy-ROCK (RNA oligomerization-enabled cryo-EM via installing kissing loops)-involves installing kissing-loop sequences onto the functionally nonessential stems of RNAs for homomeric self-assembly into closed rings with multiplied molecular weights and mitigated structural flexibility. ROCK enables cryo-EM reconstruction of the Tetrahymena group I intron at 2.98-Å resolution overall (2.85 Å for the core), allowing de novo model building of the complete RNA, including the previously unknown peripheral domains. ROCK is further applied to two smaller RNAs-the Azoarcus group I intron and the FMN riboswitch, revealing the conformational change of the former and the bound ligand in the latter. ROCK holds promise to greatly facilitate the use of cryo-EM in RNA structural studies.
External linksNat Methods / PubMed:35501384
MethodsEM (single particle)
Resolution2.85 - 5.9 Å
Structure data

24281

7r6l

EMDB-24281, PDB-7r6l:
5 prime exon-free pre-2S intermediate of the Tetrahymena group I intron, symmetry-expanded monomer from a synthetic dimeric construct
Method: EM (single particle) / Resolution: 2.85 Å

24282

7r6m

EMDB-24282, PDB-7r6m:
Post-2S intermediate of the Tetrahymena group I intron, symmetry-expanded monomer from a synthetic dimeric construct
Method: EM (single particle) / Resolution: 3.68 Å

24283

7r6n

EMDB-24283, PDB-7r6n:
Exon-free state of the Tetrahymena group I intron, symmetry-expanded monomer from a synthetic trimeric construct
Method: EM (single particle) / Resolution: 4.1 Å

EMDB-24284: Azoarcus group I intron, symmetry-expanded monomer from a synthetic trimeric construct
Method: EM (single particle) / Resolution: 4.9 Å

EMDB-24285: Fusobacterium nucleatum FMN riboswitch bound to FMN, synthetic trimeric construct
Method: EM (single particle) / Resolution: 5.9 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • tetrahymena thermophila (eukaryote)
  • Azoarcus olearius (bacteria)
  • Fusobacterium nucleatum (bacteria)
KeywordsRNA / Ribozyme / Group I intron / catalytic RNA

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