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Visualizing group II intron dynamics between the first and second steps of splicing
Journal article   Open access   Peer reviewed

Visualizing group II intron dynamics between the first and second steps of splicing

Jacopo Manigrasso, Isabel Chillón, Vito Genna, Pietro Vidossich, Srinivas Somarowthu, Anna Marie Pyle, Marco De Vivo, Marco Marcia and Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Nature communications, v 11(1), pp 2837-2837
05 Jun 2020
PMID: 32503992
url
https://doi.org/10.1038/s41467-020-16741-4View
Published, Version of Record (VoR)CC BY V4.0 Open

Abstract

Bacillaceae - genetics Catalytic Domain - genetics Crystallography, X-Ray Introns - genetics Molecular Dynamics Simulation Mutagenesis Nucleic Acid Conformation RNA Precursors - genetics RNA Precursors - metabolism RNA Splicing RNA, Bacterial - genetics RNA, Bacterial - metabolism Spatio-Temporal Analysis Spliceosomes - metabolism
Group II introns are ubiquitous self-splicing ribozymes and retrotransposable elements evolutionarily and chemically related to the eukaryotic spliceosome, with potential applications as gene-editing tools. Recent biochemical and structural data have captured the intron in multiple conformations at different stages of catalysis. Here, we employ enzymatic assays, X-ray crystallography, and molecular simulations to resolve the spatiotemporal location and function of conformational changes occurring between the first and the second step of splicing. We show that the first residue of the highly-conserved catalytic triad is protonated upon 5'-splice-site scission, promoting a reversible structural rearrangement of the active site (toggling). Protonation and active site dynamics induced by the first step of splicing facilitate the progression to the second step. Our insights into the mechanism of group II intron splicing parallels functional data on the spliceosome, thus reinforcing the notion that these evolutionarily-related molecular machines share the same enzymatic strategy.

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Web of Science research areas
Biochemistry & Molecular Biology
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