Sub-3-Å cryo-EM structure of RNA enabled by engineered homomeric self-assembly
By Di Liu, F. Thélot, J. Piccirilli, Maofu Liao, Peng Yin. 2021
43 citations recorded by the source corpus.
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. ROCK (RNA oligomerization-enabled cryo-EM via installing kissing loops) enables improved single-particle cryo-EM of RNAs. ROCK was used to generate high-quality structures of three diverse RNAs, including the Tetrahymena group I intron.
Subfields: Molecular Biology, Structural Biology
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Citations in the corpus
- Accurate RNA 3D structure prediction using a language model-based deep learning approach (2022, neutral): “In many cases, obtaining high-resolution structural information can enable a more predictive understanding of the RNA molecules of interest 4,6 .”
- Cryo-EM reveals an entangled kinetic trap in the folding of a catalytic RNA (2022, methodology): “Recently, complete structures of apo and holo TET ribozymes in the N state were solved by cryo-EM (12, 14).”