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Catalytically active Argonaute nuclease from Synechococcus elongatus

Olina, A. and Kudinova, A. and Petrova, M. and Aravin, A. and Kulbachinskiy, A. and Esyunina, D. (2019) Catalytically active Argonaute nuclease from Synechococcus elongatus. FEBS Open Bio, 9 (S1). p. 160. ISSN 2211-5463. https://resolver.caltech.edu/CaltechAUTHORS:20191017-152648736

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Abstract

Argonaute proteins, which are found in almost all eukaryotes and in many prokaryotes, use small nucleic acid guides for the recognition and cleavage of complementary nucleic acids. While the role of eukaryotic Argonautes in RNA interference is well understood, the functions of prokaryotic Argonautes remain largely unknown. It was proposed that they may provide defense against invading nucleic acids, preferably acting on DNA targets. In this work, we studied the SynAgo protein from the cyanobacterium Synechococcus elongatus. We expressed affinity-tagged SynAgo in S. elongatus, purified the protein, and sequenced and mapped associated nucleic acids. We showed that SynAgo is bound with ~18 nt small DNAs coming from all genomic regions with no obvious gene specificity. Mass-spectrometry of copurified proteins from S. elongatus also revealed several possible protein partners of SynAgo. Biochemical analysis demonstrated that SynAgo is an active nuclease that can cleave both target DNA and RNA with varying efficiency, depending on the reaction conditions and the presence of mismatches between the guide and target strands. Finally, we introduced the SynAgo gene in the E. coli genome and tested its effects on plasmid maintenance and phage infections.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/2211-5463.12675DOIAbstracts
ORCID:
AuthorORCID
Kulbachinskiy, A.0000-0002-2292-6424
Additional Information:© 2019 The Authors. FEBS Open Bio © 2019 FEBS. Issue Online: 04 July 2019. This work was supported by the grant 182907086 of the Russian Foundation for Basic Research.
Funders:
Funding AgencyGrant Number
Russian Foundation for Basic Research (RFBR)18-29-07086
Issue or Number:S1
Record Number:CaltechAUTHORS:20191017-152648736
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20191017-152648736
Official Citation:(2019), Posters. FEBS Open Bio, 9: 65-431. doi:10.1002/2211-5463.12675
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:99340
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:19 Oct 2019 04:36
Last Modified:09 Mar 2020 13:18

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