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Foreign DNA capture during CRISPR-Cas adaptive immunity.

<div class='caption-title'>Overall architecture and active site positioning of 3′–OH nucleophile</div>
<div class='caption-title'>Coordination of protospacer DNA within the complex</div>
<div class='caption-title'>Mechanism of protospacer DNA end separation</div>
Nuñez JK, Harrington LB, Kranzusch PJ, Engelman AN, Doudna JA
OpenAccess logo Nature 527 535-8 (2015)
Related entries: 5ds4, 5ds5

Cited: 109 times
EuropePMC logo PMID: 26503043

Abstract

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 segments. The universally conserved Cas1-Cas2 integrase complex catalyses spacer acquisition using a direct nucleophilic integration mechanism similar to retroviral integrases and transposases. 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 vivo. 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.

Articles - 5ds6 mentioned but not cited (1)

  1. Foreign DNA capture during CRISPR-Cas adaptive immunity. Nuñez JK, Harrington LB, Kranzusch PJ, Engelman AN, Doudna JA. Nature 527 535-538 (2015)


Reviews citing this publication (23)

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