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RNA activation-independent DNA targeting of the Type III CRISPR-Cas system by a Csm complex
KTH, School of Technology and Health (STH), Basic Science and Biomedicine, Structural Biotechnology.
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2017 (English)In: EMBO Reports, ISSN 1469-221X, E-ISSN 1469-3178, Vol. 18, no 5, p. 826-840Article in journal (Refereed) Published
Abstract [en]

The CRISPR-Cas system is an adaptive and heritable immune response that destroys invading foreign nucleic acids. The effector complex of the Type III CRISPR-Cas system targets RNA and DNA in a transcription-coupled manner, but the exact mechanism of DNA targeting by this complex remains elusive. In this study, an effector Csm holocomplex derived from Thermococcus onnurineus is reconstituted with a minimalistic combination of Csm1(1)2(1)3(3)4(1)5(1), and shows RNA targeting and RNA-activated single-stranded DNA (ssDNA) targeting activities. Unexpectedly, in the absence of an RNA transcript, it cleaves ssDNA containing a sequence complementary to the bound crRNA guide region in a manner dependent on the HD domain of the Csm1 subunit. This nuclease activity is blocked by a repeat tag found in the host CRISPR loci. The specific cleavage of ssDNA without a target RNA suggests a novel ssDNA targeting mechanism of the Type III system, which could facilitate the efficient and complete degradation of foreign nucleic acids.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2017. Vol. 18, no 5, p. 826-840
Keyword [en]
CRISPR, Csm complex, DNase, RNase, Thermococcus onnurineus
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-207890DOI: 10.15252/embr.201643700ISI: 000400446100016PubMedID: 28364023Scopus ID: 2-s2.0-85017406426OAI: oai:DiVA.org:kth-207890DiVA, id: diva2:1103509
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QC 20170530

Available from: 2017-05-30 Created: 2017-05-30 Last updated: 2017-05-30Bibliographically approved

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Hebert, Hans

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CiteExportLink to record
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