The parABSm system is involved in megaplasmid partitioning and genome integrity maintenance in Thermus thermophilus

Abstract

Abstract The characteristics of the parABS system in polyploid bacteria are barely understood. We initially analyzed the physiological functions and mechanisms of the megaplasmid parABSm system in the thermophilic polyploid bacterium Thermus thermophilus. Deletion of parABm was possible only when a plasmid-born copy of parABm was provided, indicating that these genes are conditionally essential. The cell morphology of the parABm deletion mutant (ΔparABm) was changed to some extent, and in certain extra-large or twisted cells, the nucleoids were dispersed and damaged. Compared with that of the wild type, the frequency of anucleate cells was significantly increased. Genome content analyses showed that ΔparABm had lost ∼160 kb of megaplasmid and ∼23 kb of chromosomal sequences, respectively. Genome fluorescent tagging and PFGE experiments demonstrated that the truncated megaplasmid was frequently interlinked and could not be segregated correctly; thus, certain daughter cells eventually lost the entire megaplasmid and became twisted or enlarged with damaged nucleoids. Further, we found that when the megaplasmid was lost in these cells, the toxins encoded by the megaplasmid toxin–antitoxin (TA) systems (VapBC64_65 and VapBC142_143) would exert detrimental effects, such as to fragment DNA. Thus, parABSm might ensure the existence of these TA systems, thereby preventing genomic degradation. Together, our results suggested that in T. thermophilus, the megaplasmid-encoded parABS system plays an essential role in the megaplasmid partitioning process; also it is an important determination factor for the genome integrity maintenance.