Different genome stability proteins underpin primed and naïve adaptation in E. coli CRISPR-Cas immunity
CRISPR-Cas is a prokaryotic immune system built from capture and integration of invader DNA into CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) loci, termed ‘Adaptation’, which is dependent on Cas1 and Cas2 proteins. In Escherichia coli, Cascade-Cas3 degrades invader DNA to effec...
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| Format: | Article |
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Oxford University Press
2015
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| Online Access: | https://eprints.nottingham.ac.uk/31310/ |
| _version_ | 1848794174207295488 |
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| author | Ivančić-Baće, Ivana Cass, Simon Wearne, Stephen J. Bolt, Edward L. |
| author_facet | Ivančić-Baće, Ivana Cass, Simon Wearne, Stephen J. Bolt, Edward L. |
| author_sort | Ivančić-Baće, Ivana |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | CRISPR-Cas is a prokaryotic immune system built from capture and integration of invader DNA into CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) loci, termed ‘Adaptation’, which is dependent on Cas1 and Cas2 proteins. In Escherichia coli, Cascade-Cas3 degrades invader DNA to effect immunity, termed ‘Interference’. Adaptation can interact with interference (‘primed’), or is independent of it (‘naïve’). We demonstrate that primed adaptation requires the RecG helicase and PriA protein to be present. Genetic analysis of mutant phenotypes suggests that RecG is needed to dissipate R-loops at blocked replication forks. Additionally, we identify that DNA polymerase I is important for both primed and naive adaptation, and that RecB is needed for naïve adaptation. Purified Cas1-Cas2 protein shows specificity for binding to and nicking forked DNA within single strand gaps, and collapsing forks into DNA duplexes. The data suggest that different genome stability systems interact with primed or naïve adaptation when responding to blocked or collapsed invader DNA replication. In this model, RecG and Cas3 proteins respond to invader DNA replication forks that are blocked by Cascade interference, enabling DNA capture. RecBCD targets DNA ends at collapsed forks, enabling DNA capture without interference. DNA polymerase I is proposed to fill DNA gaps during spacer integration. |
| first_indexed | 2025-11-14T19:12:00Z |
| format | Article |
| id | nottingham-31310 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:12:00Z |
| publishDate | 2015 |
| publisher | Oxford University Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-313102020-05-04T17:22:40Z https://eprints.nottingham.ac.uk/31310/ Different genome stability proteins underpin primed and naïve adaptation in E. coli CRISPR-Cas immunity Ivančić-Baće, Ivana Cass, Simon Wearne, Stephen J. Bolt, Edward L. CRISPR-Cas is a prokaryotic immune system built from capture and integration of invader DNA into CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) loci, termed ‘Adaptation’, which is dependent on Cas1 and Cas2 proteins. In Escherichia coli, Cascade-Cas3 degrades invader DNA to effect immunity, termed ‘Interference’. Adaptation can interact with interference (‘primed’), or is independent of it (‘naïve’). We demonstrate that primed adaptation requires the RecG helicase and PriA protein to be present. Genetic analysis of mutant phenotypes suggests that RecG is needed to dissipate R-loops at blocked replication forks. Additionally, we identify that DNA polymerase I is important for both primed and naive adaptation, and that RecB is needed for naïve adaptation. Purified Cas1-Cas2 protein shows specificity for binding to and nicking forked DNA within single strand gaps, and collapsing forks into DNA duplexes. The data suggest that different genome stability systems interact with primed or naïve adaptation when responding to blocked or collapsed invader DNA replication. In this model, RecG and Cas3 proteins respond to invader DNA replication forks that are blocked by Cascade interference, enabling DNA capture. RecBCD targets DNA ends at collapsed forks, enabling DNA capture without interference. DNA polymerase I is proposed to fill DNA gaps during spacer integration. Oxford University Press 2015-11-17 Article PeerReviewed Ivančić-Baće, Ivana, Cass, Simon, Wearne, Stephen J. and Bolt, Edward L. (2015) Different genome stability proteins underpin primed and naïve adaptation in E. coli CRISPR-Cas immunity. Nucleic Acids Research . pp. 1-10. ISSN 1362-4962 http://nar.oxfordjournals.org/content/43/22/10821.short doi:10.1093/nar/gkv1213 doi:10.1093/nar/gkv1213 |
| spellingShingle | Ivančić-Baće, Ivana Cass, Simon Wearne, Stephen J. Bolt, Edward L. Different genome stability proteins underpin primed and naïve adaptation in E. coli CRISPR-Cas immunity |
| title | Different genome stability proteins underpin primed
and naïve adaptation in E. coli CRISPR-Cas immunity |
| title_full | Different genome stability proteins underpin primed
and naïve adaptation in E. coli CRISPR-Cas immunity |
| title_fullStr | Different genome stability proteins underpin primed
and naïve adaptation in E. coli CRISPR-Cas immunity |
| title_full_unstemmed | Different genome stability proteins underpin primed
and naïve adaptation in E. coli CRISPR-Cas immunity |
| title_short | Different genome stability proteins underpin primed
and naïve adaptation in E. coli CRISPR-Cas immunity |
| title_sort | different genome stability proteins underpin primed
and naïve adaptation in e. coli crispr-cas immunity |
| url | https://eprints.nottingham.ac.uk/31310/ https://eprints.nottingham.ac.uk/31310/ https://eprints.nottingham.ac.uk/31310/ |