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• W1885752046 abstract "The structure of the Cas1–Cas2 complex bound to a protospacer sequence illustrates how foreign DNA is captured and measured by bacterial proteins in preparation for integration into CRISPR loci. The CRISPR–Cas system widely used for genome editing in a variety of organisms derives from a bacterial immune system. In bacteria, DNA sequences from invading phage are incorporated into the host genome in loci known as CRISPR. How these 'spacer' sequences of 30–40 base pairs are selected and generated was unclear. Jennifer Doudna and colleagues now describe the structure of the Cas1–Cas2 complex bound to a protospacer sequence. The structure illustrates how the foreign DNA is captured and cleaved by the host proteins in preparation for integration into CRISPR loci. Bacteria and archaea generate adaptive immunity against phages and plasmids by integrating foreign DNA of specific 30–40-base-pair lengths into clustered regularly interspaced short palindromic repeat (CRISPR) loci as spacer segments1,2,3,4,5,6. The universally conserved Cas1–Cas2 integrase complex catalyses spacer acquisition using a direct nucleophilic integration mechanism similar to retroviral integrases and transposases7,8,9,10,11,12,13. How the Cas1–Cas2 complex selects foreign DNA substrates for integration remains unknown. Here we present X-ray crystal structures of the Escherichia coli Cas1–Cas2 complex bound to cognate 33-nucleotide protospacer DNA substrates. The protein complex creates a curved binding surface spanning the length of the DNA and splays the ends of the protospacer to allow each terminal nucleophilic 3′-OH to enter a channel leading into the Cas1 active sites. Phosphodiester backbone interactions between the protospacer and the proteins explain the sequence-nonspecific substrate selection observed in vivo2,3,4. Our results uncover the structural basis for foreign DNA capture and the mechanism by which Cas1–Cas2 functions as a molecular ruler to dictate the sequence architecture of CRISPR loci." @default.
• W1885752046 created "2016-06-24" @default.
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• W1885752046 date "2015-10-21" @default.
• W1885752046 modified "2023-10-16" @default.
• W1885752046 title "Foreign DNA capture during CRISPR–Cas adaptive immunity" @default.
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• W1885752046 doi "https://doi.org/10.1038/nature15760" @default.
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