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Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition

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

Authors

Clare Rollie

Stefanie Schneider

Anna Sophie Brinkmann

ED BOLT ED.BOLT@NOTTINGHAM.AC.UK
Associate Professor

Malcolm F. White



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, 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 http://eprints.nottingham.ac.uk/id/eprint/31325
Publisher URL http://elifesciences.org/content/early/2015/08/17/eLife.08716
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Rollie et al Cas1 2015 eLife.pdf (9.6 Mb)
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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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