Structural and functional diversity of toxin-antitoxin-chaperone systems

Toxin-antitoxin-chaperone (TAC) systems are three-part gene clusters encoding a toxin, antitoxin, and specialized SecB-like chaperone (SecB^TA) with emerging roles in phage defense. To identify and classify SecB homologs and associated TACs across bacteria, we surveyed the full RefSeq database. Phylogenetic and gene neighborhood analyses reveal three major SecB subfamilies: two housekeeping groups and a diverse SecB^TA clade associated with eight TAC classes, five of which were previously unknown. Despite broad sequence divergence, structural predictions show conserved SecB tetrameric folds and toxin-antitoxin interfaces. The SecB chaperone phylogeny is incongruent with the identity of the TA component, suggesting modular shuffling during TAC evolution. We demonstrate toxicity of class 2 ART toxins from Escherichia coli, Bacillus subtilis, and Streptococcus gordonii, all of which we show inhibit protein synthesis. All TAC classes can be prophage encoded, indicative of phage-driven mobility and rapid diversification.