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Title | Sub-3-Å cryo-EM structure of RNA enabled by engineered homomeric self-assembly. |
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Journal, issue, pages | Nat Methods, Vol. 19, Issue 5, Page 576-585, Year 2022 |
Publish date | May 2, 2022 |
Authors | Di Liu / François A Thélot / Joseph A Piccirilli / Maofu Liao / Peng Yin / |
PubMed Abstract | 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 ...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 links | Nat Methods / PubMed:35501384 |
Methods | EM (single particle) |
Resolution | 2.85 - 5.9 Å |
Structure data | EMDB-24281, PDB-7r6l: EMDB-24282, PDB-7r6m: EMDB-24283, PDB-7r6n: EMDB-24284: Azoarcus group I intron, symmetry-expanded monomer from a synthetic trimeric construct EMDB-24285: Fusobacterium nucleatum FMN riboswitch bound to FMN, synthetic trimeric construct |
Chemicals | ChemComp-MG: |
Source |
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Keywords | RNA / Ribozyme / Group I intron / catalytic RNA |