Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition

Rollie, Clare and Schneider, Stefanie and Brinkmann, Anna Sophie and Bolt, Edward L. and White, Malcolm F. (2015) Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition. eLife . ISSN 2050-084X

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Abstract

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.

Item Type: Article
Schools/Departments: University of Nottingham UK Campus > Faculty of Medicine and Health Sciences
University of Nottingham UK Campus > Faculty of Medicine and Health Sciences > School of Life Sciences
Identification Number: https://doi.org/10.7554/eLife.08716
Depositing User: Bolt, Ed
Date Deposited: 22 Jan 2016 09:21
Last Modified: 15 Sep 2016 21:45
URI: http://eprints.nottingham.ac.uk/id/eprint/31325

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