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

Author:

Manigrasso JacopoORCID,Chillón IsabelORCID,Genna Vito,Vidossich Pietro,Somarowthu Srinivas,Pyle Anna Marie,De Vivo MarcoORCID,Marcia MarcoORCID

Abstract

AbstractGroup 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.

Funder

Howard Hughes Medical Institute

Associazione Italiana per la Ricerca sul Cancro

European Molecular Biology Laboratory

Agence Nationale de la Recherche

Agence Nationale de Recherches sur le Sida et les Hépatites Virales

Fondation ARC pour la Recherche sur le Cancer

Alliance nationale pour les sciences de la vie et de la santé (Aviesan) - ITMO Cancer: Instituts thématiques multiorganismes

Publisher

Springer Science and Business Media LLC

Subject

General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry

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