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Genome expansion by a CRISPR trimmer-integrase
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Genome expansion by a CRISPR trimmer-integrase
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Journal Article
Genome expansion by a CRISPR trimmer-integrase
2023
Overview
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.
CRISPR systems lacking Cas4 can use fused or recruited exonucleases for faithful acquisition of new CRISPR immune sequences.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 82
/ CRISPR-Associated Proteins - chemistry
/ CRISPR-Associated Proteins - metabolism
/ CRISPR-Associated Proteins - ultrastructure
/ CRISPR-Cas Systems - genetics
/ CRISPR-Cas Systems - immunology
/ Exonucleases - ultrastructure
/ Genome, Bacterial - genetics
/ Humanities and Social Sciences
/ Science
