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dc.contributor.authorRollie, Clare
dc.contributor.authorGraham, Shirley
dc.contributor.authorRouillon, Christophe
dc.contributor.authorWhite, Malcolm F.
dc.date.accessioned2017-12-08T15:30:10Z
dc.date.available2017-12-08T15:30:10Z
dc.date.issued2018-02-16
dc.identifier.citationRollie , 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/gkx1232en
dc.identifier.issn0305-1048
dc.identifier.otherPURE: 251635030
dc.identifier.otherPURE UUID: cb47f4cc-c595-4f54-bdcd-cb5cda9573e8
dc.identifier.otherWOS: 000425294400007
dc.identifier.otherWOS: 000425294400007
dc.identifier.otherScopus: 85044551490
dc.identifier.otherORCID: /0000-0003-1543-9342/work/47136084
dc.identifier.urihttp://hdl.handle.net/10023/12301
dc.descriptionThis work was supported by a grant from the Biotechnology and Biological Sciences Research Council (REF: BB/M021017/1 to MFW).en
dc.description.abstractThe 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.extent14
dc.language.isoeng
dc.relation.ispartofNucleic Acids Researchen
dc.rights© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org / licenses / by / 4.0 / ), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.subjectQH301 Biologyen
dc.subjectNDASen
dc.subjectBDCen
dc.subjectR2Cen
dc.subject.lccQH301en
dc.titlePrespacer processing and specific integration in a Type I-A CRISPR systemen
dc.typeJournal articleen
dc.contributor.sponsorBBSRCen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. School of Biologyen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.identifier.doihttps://doi.org/10.1093/nar/gkx1232
dc.description.statusPeer revieweden
dc.identifier.grantnumberBB/M021017/1en


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