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Cyanobacterial Argonautes and Cas4 family nucleases cooperate to interfere with invading DNA

Bobadilla Ugarte, Pilar ; Halter, Stefanie ; Mutte, Sumanth K ; Heijstek, Clint ; Niault, Theophile LU ; Terenin, Ilya LU orcid ; Barendse, Patrick ; Koopal, Balwina ; Roosjen, Mark and Boeren, Sjef , et al. (2025) In Molecular Cell
Abstract

Prokaryotic Argonaute proteins (pAgos) from the long-A clade are stand-alone immune systems that use small interfering DNA (siDNA) guides to recognize and cleave invading plasmid and virus DNA. Certain long-A pAgos are co-encoded with accessory proteins with unknown functions. Here, we show that cyanobacterial long-A pAgos act in conjunction with Argonaute-associated Cas4 family enzyme 1 (ACE1). Structural and biochemical analyses reveal that ACE1-associated pAgos mediate siDNA-guided DNA interference, akin to stand-alone pAgos. ACE1 is structurally homologous to the nuclease domain of bacterial DNA repair complexes and acts as a single-stranded DNA endonuclease that processes siDNA guides. pAgo and ACE1 form a heterodimeric long-A... (More)

Prokaryotic Argonaute proteins (pAgos) from the long-A clade are stand-alone immune systems that use small interfering DNA (siDNA) guides to recognize and cleave invading plasmid and virus DNA. Certain long-A pAgos are co-encoded with accessory proteins with unknown functions. Here, we show that cyanobacterial long-A pAgos act in conjunction with Argonaute-associated Cas4 family enzyme 1 (ACE1). Structural and biochemical analyses reveal that ACE1-associated pAgos mediate siDNA-guided DNA interference, akin to stand-alone pAgos. ACE1 is structurally homologous to the nuclease domain of bacterial DNA repair complexes and acts as a single-stranded DNA endonuclease that processes siDNA guides. pAgo and ACE1 form a heterodimeric long-A pAgo-ACE1 (APACE1) complex, which modulates ACE1 activity. Although ACE1-associated pAgos alone interfere with plasmids and bacteriophages, plasmid interference is boosted when pAgo and ACE1 are co-expressed. Our study reveals that pAgo-mediated immunity is enhanced by accessory proteins and broadens our mechanistic understanding of how pAgo systems interfere with invading DNA.

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Contribution to journal
publication status
epub
subject
in
Molecular Cell
publisher
Cell Press
external identifiers
  • pmid:40288374
  • scopus:105003586701
ISSN
1097-4164
DOI
10.1016/j.molcel.2025.03.025
language
English
LU publication?
yes
additional info
Copyright © 2025 The Author(s). Published by Elsevier Inc. All rights reserved.
id
5553e2e1-6058-4671-9e34-ad0ccb876109
date added to LUP
2025-04-28 09:13:01
date last changed
2025-07-15 08:03:43
@article{5553e2e1-6058-4671-9e34-ad0ccb876109,
  abstract     = {{<p>Prokaryotic Argonaute proteins (pAgos) from the long-A clade are stand-alone immune systems that use small interfering DNA (siDNA) guides to recognize and cleave invading plasmid and virus DNA. Certain long-A pAgos are co-encoded with accessory proteins with unknown functions. Here, we show that cyanobacterial long-A pAgos act in conjunction with Argonaute-associated Cas4 family enzyme 1 (ACE1). Structural and biochemical analyses reveal that ACE1-associated pAgos mediate siDNA-guided DNA interference, akin to stand-alone pAgos. ACE1 is structurally homologous to the nuclease domain of bacterial DNA repair complexes and acts as a single-stranded DNA endonuclease that processes siDNA guides. pAgo and ACE1 form a heterodimeric long-A pAgo-ACE1 (APACE1) complex, which modulates ACE1 activity. Although ACE1-associated pAgos alone interfere with plasmids and bacteriophages, plasmid interference is boosted when pAgo and ACE1 are co-expressed. Our study reveals that pAgo-mediated immunity is enhanced by accessory proteins and broadens our mechanistic understanding of how pAgo systems interfere with invading DNA.</p>}},
  author       = {{Bobadilla Ugarte, Pilar and Halter, Stefanie and Mutte, Sumanth K and Heijstek, Clint and Niault, Theophile and Terenin, Ilya and Barendse, Patrick and Koopal, Balwina and Roosjen, Mark and Boeren, Sjef and Hauryliuk, Vasili and Jinek, Martin and Westphal, Adrie H and Swarts, Daan C}},
  issn         = {{1097-4164}},
  language     = {{eng}},
  month        = {{04}},
  publisher    = {{Cell Press}},
  series       = {{Molecular Cell}},
  title        = {{Cyanobacterial Argonautes and Cas4 family nucleases cooperate to interfere with invading DNA}},
  url          = {{http://dx.doi.org/10.1016/j.molcel.2025.03.025}},
  doi          = {{10.1016/j.molcel.2025.03.025}},
  year         = {{2025}},
}