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Genome expansion by a CRISPR trimmer-integrase

CRISPR–Cas adaptive immune systems capture DNA fragments from invading mobile genetic elements and integrate them into the host genome to provide a template for RNA-guided immunity(1). CRISPR systems maintain genome integrity and avoid autoimmunity by distinguishing between self and non-self, a proc...

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Autores principales: Wang, Joy Y., Tuck, Owen T., Skopintsev, Petr, Soczek, Katarzyna M., Li, Gary, Al-Shayeb, Basem, Zhou, Julia, Doudna, Jennifer A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10284694/
https://www.ncbi.nlm.nih.gov/pubmed/37316664
http://dx.doi.org/10.1038/s41586-023-06178-2
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author Wang, Joy Y.
Tuck, Owen T.
Skopintsev, Petr
Soczek, Katarzyna M.
Li, Gary
Al-Shayeb, Basem
Zhou, Julia
Doudna, Jennifer A.
author_facet Wang, Joy Y.
Tuck, Owen T.
Skopintsev, Petr
Soczek, Katarzyna M.
Li, Gary
Al-Shayeb, Basem
Zhou, Julia
Doudna, Jennifer A.
author_sort Wang, Joy Y.
collection PubMed
description CRISPR–Cas adaptive immune systems capture DNA fragments from invading mobile genetic elements and integrate them into the host genome to provide a template for RNA-guided immunity(1). CRISPR systems maintain genome integrity and avoid autoimmunity by distinguishing between self and non-self, a process for which the CRISPR/Cas1–Cas2 integrase is necessary but not sufficient(2–5). In some microorganisms, the Cas4 endonuclease assists CRISPR adaptation(6,7), but many CRISPR–Cas systems lack Cas4(8). Here we show here that an elegant alternative pathway in a type I-E system uses an internal DnaQ-like exonuclease (DEDDh) to select and process DNA for integration using the protospacer adjacent motif (PAM). The natural Cas1–Cas2/exonuclease fusion (trimmer-integrase) catalyses coordinated DNA capture, trimming and integration. Five cryo-electron microscopy structures of the CRISPR trimmer-integrase, visualized both before and during DNA integration, show how asymmetric processing generates size-defined, PAM-containing substrates. Before genome integration, the PAM sequence is released by Cas1 and cleaved by the exonuclease, marking inserted DNA as self and preventing aberrant CRISPR targeting of the host. Together, these data support a model in which CRISPR systems lacking Cas4 use fused or recruited(9,10) exonucleases for faithful acquisition of new CRISPR immune sequences.
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spelling pubmed-102846942023-06-23 Genome expansion by a CRISPR trimmer-integrase Wang, Joy Y. Tuck, Owen T. Skopintsev, Petr Soczek, Katarzyna M. Li, Gary Al-Shayeb, Basem Zhou, Julia Doudna, Jennifer A. Nature Article CRISPR–Cas adaptive immune systems capture DNA fragments from invading mobile genetic elements and integrate them into the host genome to provide a template for RNA-guided immunity(1). CRISPR systems maintain genome integrity and avoid autoimmunity by distinguishing between self and non-self, a process for which the CRISPR/Cas1–Cas2 integrase is necessary but not sufficient(2–5). In some microorganisms, the Cas4 endonuclease assists CRISPR adaptation(6,7), but many CRISPR–Cas systems lack Cas4(8). Here we show here that an elegant alternative pathway in a type I-E system uses an internal DnaQ-like exonuclease (DEDDh) to select and process DNA for integration using the protospacer adjacent motif (PAM). The natural Cas1–Cas2/exonuclease fusion (trimmer-integrase) catalyses coordinated DNA capture, trimming and integration. Five cryo-electron microscopy structures of the CRISPR trimmer-integrase, visualized both before and during DNA integration, show how asymmetric processing generates size-defined, PAM-containing substrates. Before genome integration, the PAM sequence is released by Cas1 and cleaved by the exonuclease, marking inserted DNA as self and preventing aberrant CRISPR targeting of the host. Together, these data support a model in which CRISPR systems lacking Cas4 use fused or recruited(9,10) exonucleases for faithful acquisition of new CRISPR immune sequences. Nature Publishing Group UK 2023-06-14 2023 /pmc/articles/PMC10284694/ /pubmed/37316664 http://dx.doi.org/10.1038/s41586-023-06178-2 Text en © The Author(s) 2023 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Wang, Joy Y.
Tuck, Owen T.
Skopintsev, Petr
Soczek, Katarzyna M.
Li, Gary
Al-Shayeb, Basem
Zhou, Julia
Doudna, Jennifer A.
Genome expansion by a CRISPR trimmer-integrase
title Genome expansion by a CRISPR trimmer-integrase
title_full Genome expansion by a CRISPR trimmer-integrase
title_fullStr Genome expansion by a CRISPR trimmer-integrase
title_full_unstemmed Genome expansion by a CRISPR trimmer-integrase
title_short Genome expansion by a CRISPR trimmer-integrase
title_sort genome expansion by a crispr trimmer-integrase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10284694/
https://www.ncbi.nlm.nih.gov/pubmed/37316664
http://dx.doi.org/10.1038/s41586-023-06178-2
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