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Functional activities of DNA-guided and RNA-guided bacterial Argonaute proteins

Kropocheva, E. and Oguienko, A. and Kudinova, A. and Petrova, M. and Ryazansky, S. and Aravin, A. and Kulbachinskiy, A. (2018) Functional activities of DNA-guided and RNA-guided bacterial Argonaute proteins. FEBS Open Bio, 8 (S1). Art. No. P.01-074. ISSN 2211-5463. doi:10.1002/2211-5463.12453.

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Specific targeting of nucleic acids by Argonaute (Ago) proteins lies at the heart of RNA interference. Eukaryotic Ago’s bind small RNAs and use them as guides for target RNA recognition and cleavage. Argonaute proteins are also found in bacterial and archaeal genomes where their roles remain unclear. Structural and biochemical studies of a few prokaryotic Ago proteins showed that they can function as endonucleases in vitro and may provide cell defense against foreign genetic elements in vivo. However, most prokaryotic Ago’s are predicted to lack endonuclease activity; they also often have unusual domain architectures and are associated in the same operons with putative nucleases or helicases. In this study, we focused on prokaryotic Ago’s from several cultivable bacterial species. We showed that although eukaryotic Ago’s work only with RNA, prokaryotic proteins included in our analysis can use either RNA or DNA guides to recognize DNA targets. To define the specificity of bacterial Ago’s we expressed them in Escherichia coli and analyzed associated short nucleic acids. Furthermore, we tested nucleolytic activity of three proteins in vitro using different guide and target molecules, and also measured the affinity of Ago’s to guides and targets. The slicer activity of Ago’s is known to depend on four conserved amino acid residues in the catalytic center. We showed that, in agreement with bioinformatic predictions, two of the three proteins possess the endonuclease activity. Our results indicate that bacterial Ago proteins can cleave target nucleic acids with high specificity in vitro and can function in the heterologous E. coli system in vivo. These properties likely underlie the process of DNA/RNA interference in bacterial cells and may promote horizontal transfer of the Ago genes.

Item Type:Article
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Ryazansky, S.0000-0002-3171-2997
Kulbachinskiy, A.0000-0002-2292-6424
Additional Information:© 2018 Federation of European Biochemical Societies. First Published: 05 July 2018. This work was supported by the Grant of the Ministry of Education and Science of Russian Federation 14.W03.31.0007.
Funding AgencyGrant Number
Ministry of Education and Science of the Russian Federation14.W03.31.0007
Issue or Number:S1
Record Number:CaltechAUTHORS:20180809-161421559
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:88727
Deposited By: Tony Diaz
Deposited On:09 Aug 2018 23:24
Last Modified:16 Nov 2021 00:29

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