Snapshots of the first-step self-splicing ofTetrahymenaribozyme revealed by cryo-EM
| Autor: | Xiaojing Zhang, Shanshan Li, Grigore Pintilie, Michael Z Palo, Kaiming Zhang |
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| Rok vydání: | 2023 |
| Předmět: | |
| Zdroj: | Nucleic Acids Research. 51:1317-1325 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkac1268 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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