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Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition

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rollie2015elifee08716.pdf (8.408Mb)
Date
18/09/2015
Author
Rollie, Clare Jane Catherine
Schneider, Stefanie
Brinkmann, Anna Sophie
Bolt, Edward
White, Malcolm F
Funder
BBSRC
Grant ID
BB/M000400/1
Keywords
QH301 Biology
BDC
R2C
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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.
Citation
Rollie , C J C , Schneider , S , Brinkmann , A S , Bolt , E & White , M F 2015 , ' Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition ' , eLife , vol. 4 , e08716 . https://doi.org/10.7554/eLife.08716
Publication
eLife
Status
Peer reviewed
DOI
https://doi.org/10.7554/eLife.08716
ISSN
2050-084X
Type
Journal article
Rights
Copyright © Rollie et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
Description
This work was supported by a grant from the Biotechnology and Biological Sciences Research Council (REF: BB/M000400/1 to MFW).
Collections
  • University of St Andrews Research
URI
http://hdl.handle.net/10023/7519

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