Physical and Functional Interplay between MazF1Bif and Its Noncognate Antitoxins from Bifidobacterium longum.

Bifidobacterium longum strain JDM301, a widely used commercial strain in China, encodes at least two MazEF-like modules and one RelBE-like toxin-antitoxin (TA) system in its chromosome, designated MazE1F1Bif, MazE2F2Bif, and RelBEBif, respectively. Bacterial TA systems play an important role in several stress responses, but the relationship between these TA systems is largely unknown. In this study, the interactions between MazF1Bif and MazE2Bif or RelBBif were assessed in B. longum strain JDM301. MazF1Bif caused the degradation of tufABif mRNA, and its toxicity was inhibited by forming a protein complex with its cognate antitoxin, MazE1Bif Notably, MazF1Bif toxicity was also partially neutralized when jointly expressed with noncognate antitoxin MazE2Bif or RelBBif Our results show that the two noncognate antitoxins also inhibited mRNA degradation caused by MazF1Bif toxin. Furthermore, the physical interplay between MazF1Bif and its noncognate antitoxins was confirmed by immunoprecipitation. These results suggest that MazF1Bif can arrest cell growth and that MazF1Bif toxicity can be neutralized by its cognate and noncognate antitoxins. These results imply that JDM301 uses a sophisticated toxin-antitoxin interaction network to alter its physiology when coping with environmental stress.IMPORTANCE Although toxin-antitoxin (TA) systems play an important role in several stress responses, the regulatory mechanisms of multiple TA system homologs in the bacterial genome remain largely unclear. In this study, the relationships between MazE1F1Bif and the other two TA systems of Bifidobacterium longum strain JDM301 were explored, and the interactions between MazF1Bif and MazE2Bif or RelBBif were characterized. In addition, the mRNA degradation activity of MazF1Bif was demonstrated. In particular, the interaction of the toxin with noncognate antitoxins was shown, even between different TA families (MazF1Bif toxin and RelBBif antitoxin) in JDM301. This work provides insight into the regulatory mechanisms of TA systems implicated in the stress responses of bifidobacteria.