Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition

Rollie, Clare; Schneider, Stefanie; Brinkmann, Anna Sophie; Bolt, Edward L.; White, Malcolm F.

Abstract

© Rollie et al. The adaptive prokaryotic immune system CRISPR-Cas provides RNA-mediated protection from invading genetic elements. The fundamental basis of the system is the ability to capture small pieces of foreign DNA for incorporation into the genome at the CRISPR locus, a process known as Adaptation, which is dependent on the Cas1 and Cas2 proteins. We demonstrate that Cas1 catalyses an efficient trans-esterification reaction on branched DNA substrates, which represents the reverse- or disintegration reaction. Cas1 from both Escherichia coli and Sulfolobus solfataricus display sequence specific activity, with a clear preference for the nucleotides flanking the integration site at the leader-repeat 1 boundary of the CRISPR locus. Cas2 is not required for this activity and does not influence the specificity. This suggests that the inherent sequence specificity of Cas1 is a major determinant of the adaptation process.

Citation

Rollie, C., Schneider, S., Brinkmann, A. S., Bolt, E. L., & White, M. F. (2015). Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition. eLife, 4, Article e08716. https://doi.org/10.7554/elife.08716

• Journal Article Type: Article
• Acceptance Date: Aug 17, 2015
• Online Publication Date: Aug 18, 2015
• Publication Date: Aug 18, 2015
• Deposit Date: Jan 22, 2016
• Publicly Available Date: Jan 22, 2016
• Journal: eLife
• Electronic ISSN: 2050-084X
• Publisher: eLife Sciences Publications
• Peer Reviewed: Peer Reviewed
• Volume: 4
• Article Number: e08716
• DOI: https://doi.org/10.7554/elife.08716
• Public URL: https://nottingham-repository.worktribe.com/output/758796
• Publisher URL: http://elifesciences.org/content/early/2015/08/17/eLife.08716
• Contract Date: Jan 22, 2016