Cyanobacterial Argonautes and Cas4 family nucleases cooperate to interfere with invading DNA
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.
Authors
- Martin Jínek (ORCID: https://orcid.org/0000-0002-7601-210X)
- Ilya M. Terenin (ORCID: https://orcid.org/0000-0002-4459-0781)
- Vasili Hauryliuk (ORCID: https://orcid.org/0000-0003-2389-5057)
- Sjef Boeren
- Adrie H. Westphal (ORCID: https://orcid.org/0000-0003-4264-6267)
- Théophile Niault (ORCID: https://orcid.org/0000-0001-7241-8914)
- Pilar Bobadilla Ugarte (ORCID: https://orcid.org/0000-0003-4147-167X)
- Balwina Koopal (ORCID: https://orcid.org/0000-0002-5310-7528)
- Mark Roosjen
- Sumanth Mutte (ORCID: https://orcid.org/0000-0003-3376-2354)
- Daan C. Swarts (ORCID: https://orcid.org/0000-0003-4412-9191)
- Patrick Barendse (ORCID: https://orcid.org/0000-0002-6332-354X)
- Clint Heijstek (ORCID: https://orcid.org/0009-0007-0567-664X)
- Susan A. Halter
Institutions
- Lund University (SE)
- University of Zurich (CH)
- Wageningen University & Research (NL)
Publication Details
- Journal
- Molecular Cell
- Published
- 2025-04-26
- DOI
- https://doi.org/10.1016/j.molcel.2025.03.025
- Citations
- 5
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 4.03
Funders
- European Molecular Biology Organization
- European Commission
- Technische Universiteit Delft
- Consejo Nacional de Ciencia y Tecnología
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek
- Knut och Alice Wallenbergs Stiftelse
- Paul Scherrer Institut
- Vetenskapsrådet
- Universität Zürich
- Wageningen University and Research
- H2020 European Research Council