Zander, A;
Willkomm, S;
Ofer, S;
van Wolferen, M;
Egert, L;
Buchmeier, S;
Stöckl, S;
... Grohmann, D; + view all
(2017)
Guide-independent DNA cleavage by archaeal Argonaute from Methanocaldococcus jannaschii.
Nature Microbiology
, 2
, Article 17034. 10.1038/nmicrobiol.2017.34.
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Abstract
Prokaryotic Argonaute proteins acquire guide strands derived from invading or mobile genetic elements, via an unknown pathway, to direct guide-dependent cleavage of foreign DNA. Here, we report that Argonaute from the archaeal organism Methanocaldococcus jannaschii (MjAgo) possesses two modes of action: the canonical guide-dependent endonuclease activity and a non-guided DNA endonuclease activity. The latter allows MjAgo to process long double-stranded DNAs, including circular plasmid DNAs and genomic DNAs. Degradation of substrates in a guide-independent fashion primes MjAgo for subsequent rounds of DNA cleavage. Chromatinized genomic DNA is resistant to MjAgo degradation, and recombinant histones protect DNA from cleavage in vitro. Mutational analysis shows that key residues important for guide-dependent target processing are also involved in guide-independent MjAgo function. This is the first characterization of guide-independent cleavage activity for an Argonaute protein potentially serving as a guide biogenesis pathway in a prokaryotic system.
| Type: | Article |
|---|---|
| Title: | Guide-independent DNA cleavage by archaeal Argonaute from Methanocaldococcus jannaschii |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1038/nmicrobiol.2017.34 |
| Publisher version: | http://doi.org/10.1038/nmicrobiol.2017.34 |
| Language: | English |
| Additional information: | © 2017 Macmillan Publishers Limited, part of Springer Nature. All rights reserved. This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1546543 |
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