Snapshots of the second-step self-splicing of Tetrahymena ribozyme revealed by cryo-EM

Shanshan Li (), Michael Z. Palo, Xiaojing Zhang, Grigore Pintilie and Kaiming Zhang ()
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Shanshan Li: University of Science and Technology of China
Michael Z. Palo: Stanford University
Xiaojing Zhang: University of Science and Technology of China
Grigore Pintilie: Stanford University
Kaiming Zhang: University of Science and Technology of China

Nature Communications, 2023, vol. 14, issue 1, 1-10

Abstract: Abstract Group I introns are catalytic RNAs that coordinate two consecutive transesterification reactions for self-splicing. To understand how the group I intron promotes catalysis and coordinates self-splicing reactions, we determine the structures of L-16 Tetrahymena ribozyme in complex with a 5′-splice site analog product and a 3′-splice site analog substrate using cryo-EM. We solve six conformations from a single specimen, corresponding to different splicing intermediates after the first ester-transfer reaction. The structures reveal dynamics during self-splicing, including large conformational changes of the internal guide sequence and the J5/4 junction as well as subtle rearrangements of active-site metals and the hydrogen bond formed between the 2′-OH group of A261 and the N2 group of guanosine substrate. These results help complete a detailed structural and mechanistic view of this paradigmatic group I intron undergoing the second step of self-splicing.

Date: 2023
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DOI: 10.1038/s41467-023-36724-5

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