Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition
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...
| Main Authors: | , , , , |
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| Format: | Article |
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eLife Sciences Publications
2015
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| Online Access: | https://eprints.nottingham.ac.uk/31325/ |
| _version_ | 1848794177523941376 |
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| author | Rollie, Clare Schneider, Stefanie Brinkmann, Anna Sophie Bolt, Edward L. White, Malcolm F. |
| author_facet | Rollie, Clare Schneider, Stefanie Brinkmann, Anna Sophie Bolt, Edward L. White, Malcolm F. |
| author_sort | Rollie, Clare |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:12:03Z |
| format | Article |
| id | nottingham-31325 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:12:03Z |
| publishDate | 2015 |
| publisher | eLife Sciences Publications |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-313252020-05-04T17:14:57Z https://eprints.nottingham.ac.uk/31325/ Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition Rollie, Clare Schneider, Stefanie Brinkmann, Anna Sophie Bolt, Edward L. White, Malcolm F. 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. eLife Sciences Publications 2015-08-18 Article PeerReviewed Rollie, Clare, Schneider, Stefanie, Brinkmann, Anna Sophie, Bolt, Edward L. and White, Malcolm F. (2015) Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition. eLife . ISSN 2050-084X http://elifesciences.org/content/early/2015/08/17/eLife.08716 doi:10.7554/eLife.08716 doi:10.7554/eLife.08716 |
| spellingShingle | Rollie, Clare Schneider, Stefanie Brinkmann, Anna Sophie Bolt, Edward L. White, Malcolm F. Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition |
| title | Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition |
| title_full | Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition |
| title_fullStr | Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition |
| title_full_unstemmed | Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition |
| title_short | Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition |
| title_sort | intrinsic sequence specificity of the cas1 integrase directs new spacer acquisition |
| url | https://eprints.nottingham.ac.uk/31325/ https://eprints.nottingham.ac.uk/31325/ https://eprints.nottingham.ac.uk/31325/ |