A bacterial CARD–NLR-like immune system controls the release of gene transfer agents

Bacteria use immune systems to detect and defend against mobile genetic elements including phages. Gene transfer agents (GTAs) are domesticated prophages with phage-like characteristics including the ability to induce host cell lysis for gene transfer. Whether GTAs elicit or avoid bacterial immune systems is poorly understood. Here, a transposon mutagenesis with deep sequencing screen in Caulobacter crescentus identified a tripartite system, LypABC, essential for GTA-mediated cell lysis and gene transfer. LypABC resembles a caspase recruitment domain–nucleotide-binding leucine-rich repeat (CARD–NLR) anti-phage defence system. LypABC is dispensable for DNA packaging into GTA particles but required for host cell lysis, involving the peptidase domains of LypA and LypC, and the ATPase domain of LypB. As LypABC overproduction is toxic, strict regulation through the transcriptional repressor CdxB is required. CdxB binds the promoters of lypABC and of essential GTA activator genes, coupling GTA activation to host cell lysis. Our findings suggest that bacterial immune systems can be co-opted to support horizontal gene transfer by GTAs.