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dc.contributor.authorSokolowski, Richard David
dc.contributor.authorGraham, Shirley
dc.contributor.authorWhite, Malcolm F
dc.date.accessioned2014-04-22T13:31:02Z
dc.date.available2014-04-22T13:31:02Z
dc.date.issued2014-06-02
dc.identifier.citationSokolowski , R D , Graham , S & White , M F 2014 , ' Cas6 specificity and CRISPR RNA loading in a complex CRISPR-Cas system ' Nucleic Acids Research , vol. 42 , no. 10 , pp. 6532-6541 . DOI: 10.1093/nar/gku308en
dc.identifier.issn0305-1048
dc.identifier.otherPURE: 112703281
dc.identifier.otherPURE UUID: 78fd5bb8-8ae8-4509-b2cf-a36a0b396a38
dc.identifier.otherWOS: 000338768100041
dc.identifier.otherScopus: 84903202938
dc.identifier.urihttp://hdl.handle.net/10023/4582
dc.descriptionThis research was funded in part by Biotechnology and Biological Sciences Research Council [BB/K000314/1]. The APC was paid through RCUK OA block grant funds.en
dc.description.abstractCRISPR-Cas is an adaptive prokaryotic immune system, providing protection against viruses and other mobile genetic elements. In type I and type III CRISPR-Cas systems, CRISPR RNA (crRNA) is generated by cleavage of a primary transcript by the Cas6 endonuclease and loaded into multisubunit surveillance/effector complexes, allowing homology-directed detection and cleavage of invading elements. Highly studied CRISPR-Cas systems such as those in Escherichia coli and Pseudomonas aeruginosa have a single Cas6 enzyme that is an integral subunit of the surveillance complex. By contrast, Sulfolobus solfataricus has a complex CRISPR-Cas system with three types of surveillance complexes (Cascade/type I-A, CSM/type III-A and CMR/type III-B), five Cas6 paralogues and two different CRISPR-repeat families (AB and CD). Here, we investigate the kinetic properties of two different Cas6 paralogues from S. solfataricus. The Cas6-1 subtype is specific for CD-family CRISPR repeats, generating crRNA by multiple turnover catalysis whilst Cas6-3 has a broader specificity and also processes a non-coding RNA with a CRISPR repeat-related sequence. Deep sequencing of crRNA in surveillance complexes reveals a biased distribution of spacers derived from AB and CD loci, suggesting functional coupling between Cas6 paralogues and their downstream effector complexes.en
dc.format.extent10en
dc.language.isoeng
dc.relation.ispartofNucleic Acids Researchen
dc.rights© 2014, The Authors. 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/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly citeden
dc.subjectCRISPR-Casen
dc.subjectProkaryotic immune systemen
dc.subjectCRISPR RNA (crRNA)en
dc.subjectCas6en
dc.subjectSulfolobus solfataricusen
dc.titleCas6 specificity and CRISPR RNA loading in a complex CRISPR-Cas systemen
dc.typeJournal articleen
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/gku308
dc.description.statusPeer revieweden


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