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Multiple nucleic acid cleavage modes in divergent type III CRISPR systems

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Nucl._Acids_Res._2016_Zhang_nar_gkw020.pdf (8.118Mb)
Date
29/02/2016
Author
Zhang, Jing
Graham, Shirley
Tello, Agnes
Liu, Huanting
White, Malcolm Frederick
Keywords
QH301 Biology
QD Chemistry
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Abstract
CRISPR-Cas is an RNA-guided adaptive immune system that protects bacteria and archaea from invading nucleic acids. Type III systems (Cmr, Csm) have been shown to cleave RNA targets in vitro and some are capable of transcription-dependent DNA targeting. The crenarchaeon Sulfolobus solfataricus has two divergent subtypes of the type III system (Sso-IIID and a Cmr7-containing variant of Sso-IIIB). Here, we report that both the Sso-IIID and Sso-IIIB complexes cleave cognate RNA targets with a ruler mechanism and 6 or 12 nt spacing that relates to the organization of the Cas7 backbone. This backbone-mediated cleavage activity thus appears universal for the type III systems. The Sso-IIIB complex is also known to possess a distinct ‘UA’ cleavage mode. The predominant activity observed in vitro depends on the relative molar concentration of protein and target RNA. The Sso-IIID complex can cleave plasmid DNA targets in vitro, generating linear DNA products with an activity that is dependent on both the cyclase and HD nuclease domains of the Cas10 subunit, suggesting a role for both nuclease active sites in the degradation of double-stranded DNA targets.
Citation
Zhang , J , Graham , S , Tello , A , Liu , H & White , M F 2016 , ' Multiple nucleic acid cleavage modes in divergent type III CRISPR systems ' , Nucleic Acids Research , vol. 44 , no. 4 , pp. 1789-1799 . https://doi.org/10.1093/nar/gkw020
Publication
Nucleic Acids Research
Status
Peer reviewed
DOI
https://doi.org/10.1093/nar/gkw020
ISSN
0305-1048
Type
Journal article
Rights
Copyright The Author(s) 2016. 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-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
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  • University of St Andrews Research
URI
http://hdl.handle.net/10023/8058

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