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Title | Snapshots of the first-step self-splicing of Tetrahymena ribozyme revealed by cryo-EM. |
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Journal, issue, pages | Nucleic Acids Res, Vol. 51, Issue 3, Page 1317-1325, Year 2023 |
Publish date | Feb 22, 2023 |
Authors | Xiaojing Zhang / Shanshan Li / Grigore Pintilie / Michael Z Palo / Kaiming Zhang / |
PubMed Abstract | Tetrahymena ribozyme is a group I intron, whose self-splicing is the result of two sequential ester-transfer reactions. To understand how it facilitates catalysis in the first self-splicing reaction, ...Tetrahymena ribozyme is a group I intron, whose self-splicing is the result of two sequential ester-transfer reactions. To understand how it facilitates catalysis in the first self-splicing reaction, we used cryogenic electron microscopy (cryo-EM) to resolve the structures of L-16 Tetrahymena ribozyme complexed with a 11-nucleotide 5'-splice site analog substrate. Four conformations were achieved to 4.14, 3.18, 3.09 and 2.98 Å resolutions, respectively, corresponding to different splicing intermediates during the first enzymatic reaction. Comparison of these structures reveals structural alterations, including large conformational changes in IGS/IGSext (P1-P1ext duplex) and J5/4, as well as subtle local rearrangements in the G-binding site. These structural changes are required for the enzymatic activity of the Tetrahymena ribozyme. Our study demonstrates the ability of cryo-EM to capture dynamic RNA structural changes, ushering in a new era in the analysis of RNA structure-function by cryo-EM. |
External links | Nucleic Acids Res / PubMed:36660826 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.98 - 4.14 Å |
Structure data | EMDB-33736, PDB-7yc8: EMDB-33738, PDB-7ycg: EMDB-33739, PDB-7ych: EMDB-33740, PDB-7yci: |
Chemicals | ChemComp-GTP: ChemComp-MG: |
Source |
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Keywords | RNA / Tetrahymena ribozyme / first step of self-splicing / conformation 1 / conformation 2 / conformation 3 / conformation 4 |