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Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage
CRISPR-Cas systems are an adaptive immune system in bacteria and archaea that utilize CRISPR RNA-guided surveillance complexes to target complementary RNA or DNA for destruction(1–5). Target RNA cleavage at regular intervals is characteristic of type III effector complexes; however, the mechanism ha...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Journal Experts
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10168453/ https://www.ncbi.nlm.nih.gov/pubmed/37163044 http://dx.doi.org/10.21203/rs.3.rs-2837968/v1 |
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author | Schwartz, Evan A. Bravo, Jack P.K. Ahsan, Mohd Macias, Luis A. McCafferty, Caitlyn L. Dangerfield, Tyler L. Walker, Jada N. Brodbelt, Jennifer S. Palermo, Giulia Fineran, Peter C. Fagerlund, Robert D. Taylor, David W. |
author_facet | Schwartz, Evan A. Bravo, Jack P.K. Ahsan, Mohd Macias, Luis A. McCafferty, Caitlyn L. Dangerfield, Tyler L. Walker, Jada N. Brodbelt, Jennifer S. Palermo, Giulia Fineran, Peter C. Fagerlund, Robert D. Taylor, David W. |
author_sort | Schwartz, Evan A. |
collection | PubMed |
description | CRISPR-Cas systems are an adaptive immune system in bacteria and archaea that utilize CRISPR RNA-guided surveillance complexes to target complementary RNA or DNA for destruction(1–5). Target RNA cleavage at regular intervals is characteristic of type III effector complexes; however, the mechanism has remained enigmatic(6,7). Here, we determine the structures of the Synechocystis type III-Dv complex, an evolutionary intermediate in type III effectors(8,9), in pre- and post-cleavage states, which show metal ion coordination in the active sites. Using structural, biochemical, and quantum/classical molecular dynamics simulation, we reveal the structure and dynamics of the three catalytic sites, where a 2’-OH of the ribose on the target RNA acts as a nucleophile for in line self-cleavage of the upstream scissile phosphate. Strikingly, the arrangement at the catalytic residues of most type III complexes resembles the active site of ribozymes, including the hammerhead, pistol, and Varkud satellite ribozymes. Thus, type III CRISPR-Cas complexes function as protein-assisted ribozymes, and their programmable nature has important implications for how these complexes could be repurposed for applications. |
format | Online Article Text |
id | pubmed-10168453 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Journal Experts |
record_format | MEDLINE/PubMed |
spelling | pubmed-101684532023-05-10 Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage Schwartz, Evan A. Bravo, Jack P.K. Ahsan, Mohd Macias, Luis A. McCafferty, Caitlyn L. Dangerfield, Tyler L. Walker, Jada N. Brodbelt, Jennifer S. Palermo, Giulia Fineran, Peter C. Fagerlund, Robert D. Taylor, David W. Res Sq Article CRISPR-Cas systems are an adaptive immune system in bacteria and archaea that utilize CRISPR RNA-guided surveillance complexes to target complementary RNA or DNA for destruction(1–5). Target RNA cleavage at regular intervals is characteristic of type III effector complexes; however, the mechanism has remained enigmatic(6,7). Here, we determine the structures of the Synechocystis type III-Dv complex, an evolutionary intermediate in type III effectors(8,9), in pre- and post-cleavage states, which show metal ion coordination in the active sites. Using structural, biochemical, and quantum/classical molecular dynamics simulation, we reveal the structure and dynamics of the three catalytic sites, where a 2’-OH of the ribose on the target RNA acts as a nucleophile for in line self-cleavage of the upstream scissile phosphate. Strikingly, the arrangement at the catalytic residues of most type III complexes resembles the active site of ribozymes, including the hammerhead, pistol, and Varkud satellite ribozymes. Thus, type III CRISPR-Cas complexes function as protein-assisted ribozymes, and their programmable nature has important implications for how these complexes could be repurposed for applications. American Journal Experts 2023-04-27 /pmc/articles/PMC10168453/ /pubmed/37163044 http://dx.doi.org/10.21203/rs.3.rs-2837968/v1 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use. |
spellingShingle | Article Schwartz, Evan A. Bravo, Jack P.K. Ahsan, Mohd Macias, Luis A. McCafferty, Caitlyn L. Dangerfield, Tyler L. Walker, Jada N. Brodbelt, Jennifer S. Palermo, Giulia Fineran, Peter C. Fagerlund, Robert D. Taylor, David W. Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage |
title | Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage |
title_full | Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage |
title_fullStr | Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage |
title_full_unstemmed | Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage |
title_short | Type III CRISPR-Cas complexes act as protein-assisted ribozymes during target RNA cleavage |
title_sort | type iii crispr-cas complexes act as protein-assisted ribozymes during target rna cleavage |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10168453/ https://www.ncbi.nlm.nih.gov/pubmed/37163044 http://dx.doi.org/10.21203/rs.3.rs-2837968/v1 |
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