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

Journal Article · · Nature Communications
 [1];  [2];  [3];  [1];  [4];  [5];  [1];  [2]
  1. Istituto Italiano di Tecnologia, Genoa (Italy)
  2. European Molecular Biology Lab. (EMBL), Grenoble (France)
  3. Inst. for Research in Biomedicine (IRB), Barcelona (Spain)
  4. Drexel Univ., Philadelphia, PA (United States)
  5. Cellular and Developmental Biology, New Haven, CT (United States); Howard Hughes Medical Inst., Chevy Chase, MD (United States)
+ Show Author Affiliations
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.
Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
Agence Nationale de Recherche sur le Sida et les hépatites virales (ANRS); Agence Nationale de la Recherché (ANR); Fondation Arc Pour La Recherche Sur Le Cancer (Fondation Arc); ITMO Cancer; Italian Association for Cancer Research (AIRC)
OSTI ID:
1637956
Journal Information:
Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 11; ISSN 2041-1723
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

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