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Visualizing group II intron dynamics between the first and second steps of splicing
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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7275048/ https://www.ncbi.nlm.nih.gov/pubmed/32503992 http://dx.doi.org/10.1038/s41467-020-16741-4 |
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author | Manigrasso, Jacopo Chillón, Isabel Genna, Vito Vidossich, Pietro Somarowthu, Srinivas Pyle, Anna Marie De Vivo, Marco Marcia, Marco |
author_facet | Manigrasso, Jacopo Chillón, Isabel Genna, Vito Vidossich, Pietro Somarowthu, Srinivas Pyle, Anna Marie De Vivo, Marco Marcia, Marco |
author_sort | Manigrasso, Jacopo |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7275048 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-72750482020-06-16 Visualizing group II intron dynamics between the first and second steps of splicing Manigrasso, Jacopo Chillón, Isabel Genna, Vito Vidossich, Pietro Somarowthu, Srinivas Pyle, Anna Marie De Vivo, Marco Marcia, Marco Nat Commun Article 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. Nature Publishing Group UK 2020-06-05 /pmc/articles/PMC7275048/ /pubmed/32503992 http://dx.doi.org/10.1038/s41467-020-16741-4 Text en © The Author(s) 2020, corrected publication 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Manigrasso, Jacopo Chillón, Isabel Genna, Vito Vidossich, Pietro Somarowthu, Srinivas Pyle, Anna Marie De Vivo, Marco Marcia, Marco Visualizing group II intron dynamics between the first and second steps of splicing |
title | Visualizing group II intron dynamics between the first and second steps of splicing |
title_full | Visualizing group II intron dynamics between the first and second steps of splicing |
title_fullStr | Visualizing group II intron dynamics between the first and second steps of splicing |
title_full_unstemmed | Visualizing group II intron dynamics between the first and second steps of splicing |
title_short | Visualizing group II intron dynamics between the first and second steps of splicing |
title_sort | visualizing group ii intron dynamics between the first and second steps of splicing |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7275048/ https://www.ncbi.nlm.nih.gov/pubmed/32503992 http://dx.doi.org/10.1038/s41467-020-16741-4 |
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