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Prespacer processing and specific integration in a Type I-A CRISPR system
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dc.contributor.author | Rollie, Clare | |
dc.contributor.author | Graham, Shirley | |
dc.contributor.author | Rouillon, Christophe | |
dc.contributor.author | White, Malcolm F. | |
dc.date.accessioned | 2017-12-08T15:30:10Z | |
dc.date.available | 2017-12-08T15:30:10Z | |
dc.date.issued | 2018-02-16 | |
dc.identifier | 251635030 | |
dc.identifier | cb47f4cc-c595-4f54-bdcd-cb5cda9573e8 | |
dc.identifier | 000425294400007 | |
dc.identifier | 000425294400007 | |
dc.identifier | 85044551490 | |
dc.identifier.citation | Rollie , C , Graham , S , Rouillon , C & White , M F 2018 , ' Prespacer processing and specific integration in a Type I-A CRISPR system ' , Nucleic Acids Research , vol. 46 , no. 3 , pp. 1007-1020 . https://doi.org/10.1093/nar/gkx1232 | en |
dc.identifier.issn | 0305-1048 | |
dc.identifier.other | ORCID: /0000-0003-1543-9342/work/47136084 | |
dc.identifier.uri | https://hdl.handle.net/10023/12301 | |
dc.description | This work was supported by a grant from the Biotechnology and Biological Sciences Research Council (REF: BB/M021017/1 to MFW). | en |
dc.description.abstract | The CRISPR–Cas system for prokaryotic adaptive immunity provides RNA-mediated protection from viruses and mobile genetic elements. Adaptation is dependent on the Cas1 and Cas2 proteins along with varying accessory proteins. Here we analyse the process in Sulfolobus solfataricus, showing that while Cas1 and Cas2 catalyze spacer integration in vitro, host factors are required for specificity. Specific integration also requires at least 400 bp of the leader sequence, and is dependent on the presence of hydrolysable ATP, suggestive of an active process that may involve DNA remodelling. Specific spacer integration is associated with processing of prespacer 3′ ends in a PAM-dependent manner. This is reflected in PAM-dependent processing of prespacer 3′ ends in vitro in the presence of cell lysate or the Cas4 nuclease, in a reaction consistent with PAM-directed binding and protection of prespacer DNA. These results highlight the diverse interplay between CRISPR–Cas elements and host proteins across CRISPR types. | |
dc.format.extent | 14 | |
dc.format.extent | 3922336 | |
dc.language.iso | eng | |
dc.relation.ispartof | Nucleic Acids Research | en |
dc.subject | QH301 Biology | en |
dc.subject | NDAS | en |
dc.subject | BDC | en |
dc.subject | R2C | en |
dc.subject.lcc | QH301 | en |
dc.title | Prespacer processing and specific integration in a Type I-A CRISPR system | en |
dc.type | Journal article | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.institution | University of St Andrews. School of Biology | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.identifier.doi | 10.1093/nar/gkx1232 | |
dc.description.status | Peer reviewed | en |
dc.identifier.grantnumber | BB/M021017/1 | en |
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