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Bacterial Argonaute nucleases reveal different modes of DNA targeting in vitro and in vivo
Open accessPreprint — published as 10.1093/nar/gkad290 · Paper page
Abstract
Prokaryotic Argonaute proteins (pAgos) are homologs of eukaryotic Argonautes (eAgos) that were similarly proposed to play a role in cell defense against invaders. However, pAgos are much more diverse than eAgos and very little is known about their functional activity and target specificity in vivo . Here, we describe five pAgo proteins from mesophilic bacteria that act as DNA-guided DNA endonucleases and analyze their ability to target chromosomal and invader DNA. In vitro , the analyzed proteins use small guide DNAs for precise cleavage of single-stranded DNA at a wide range of temperatures. Upon their expression in Escherichia coli , all five pAgos are loaded with small DNAs preferentially produced from plasmid DNA and from chromosomal regions of replication termination. One of the tested pAgos, EmaAgo from Exiguobacterium marinum can induce DNA interference between multicopy sequences resulting in targeted processing of homologous plasmid and chromosomal loci. EmaAgo also protects bacteria from bacteriophage infection and is preferentially loaded with phage guide DNAs suggesting that the ability of pAgos to target multicopy elements may be crucial for their protective function. The wide spectrum of pAgo activities suggests that they may have diverse functions in vivo and paves the way for their use in biotechnology.
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Metadata
- Authors
- Lisitskaya, L., Kropocheva, E., Agapov, A., Prostova, M., Panteleev, V., Yudin, D., Ryazansky, S., Kuzmenko, A., Aravin, A. A., Esyunina, D., & Kulbachinskiy, A.
- Affiliations
- Russian Academy of Sciences (RU)
- Kurchatov Institute (RU)
- California Institute of Technology (US)
- Corresponding author
- Kulbachinskiy, Andrey
- ORCID
- Venue
- Nucleic Acids Research
- Publisher
- openRxiv
- Published
- 2022-09-09
- Volume
- 51
- Issue
- 10
- Pages
- 5106-5124
- ISSN
- 0305-1048 (ISSN-L)
- PMC
- PMC10250240
- OpenAlex
- W4366816650
- Cited by
- 26
- References
- 1
- Citations by year
- 2026: 7 · 2025: 10 · 2024: 6 · 2023: 2 · 2022: 1
- FWCI
- 2.23
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by-nc
- Locations
- Nucleic Acids Research (publishedVersion, cc-by-nc)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- Ministerstwo Edukacji i Nauki (075-15-2021-1062)
- Russian Science Foundation (20-74-10127, 19-14-00359)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Immunology and Microbiology › Virology › Bacteriophages and microbial interactions
- Keywords
- bacteriophage defense, phage resistance, replication termination, Escherichia coli, single-stranded DNA cleavage, mesophilic bacteria
- MeSH terms
- Bacteria, DNA, DNA, Bacterial, Endonucleases, Plasmids, Prokaryotic Cells, Eukaryota, Argonaute Proteins
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