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

ABSTRACT: Prokaryotic Argonaute proteins (pAgos) from the long-A clade are stand-alone immune systems that use small interfering (si)DNA 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 APACE1 complex, which modulates ACE1 activity. While 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.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Escherichia Coli

SUBMITTER: Mark Roosjen

LAB HEAD: Daan Swarts

PROVIDER: PXD060946 | Pride | 2025-04-15

REPOSITORIES: Pride

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			Action	DRS
	MR3291_MaAGOACE1_1.mgf	Mgf
	MR3291_MaAGOACE1_1.raw	Raw
	MR3292_MaAGOACE1_2.mgf	Mgf
	MR3292_MaAGOACE1_2.raw	Raw
	MR3293_MaAGOACE1_3.mgf	Mgf

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