Dynamics of chromosome replication and its relationship to predatory attack lifestyles in Bdellovibrio bacteriovorus

Abstract

Bdellovibrio bacteriovorus is a small Gram-negative, an obligate predatory bacterium that is largely found in wet, aerobic environments (i.e. soil). This bacterium attacks and invades other Gram-negative bacteria, including animal and plant pathogens. The intriguing life cycle of B. bacteriovorus consists of two phases: a free-living non-replicative attack phase wherein the predatory bacterium searches for its prey, and a reproductive phase, in which B. bacteriovorus degrades a hosts macromolecules and reuses them for its own growth and chromosome replication. Although the cell biology of this predatory bacterium has gained considerable interest in recent years, we know almost nothing about the dynamics of chromosome replication in B. bacteriovorus. Here, we performed a real-time investigation into the subcellular localization of the replisome(s) in single cells of B. bacteriovorus. Our results confirm that in B. bacteriovorus chromosome replication fires only during the reproductive phase, and show for the first time that this predatory bacterium exhibits a novel spatiotemporal arrangement of chromosome replication. The replication process starts at the invasive pole of the predatory bacterium inside the prey cell and proceeds until several copies of the chromosome have been completely synthesized. This chromosome replication is not coincident with the predator-cell division, and it terminates shortly before synchronous predator-filament septation occurs. In addition, we demonstrate that if this lifecycle fails in some cells of B. bacteriovorus, they can instead use two prey cells sequentially to complete their life cycle.\n\nImportanceNew strategies are needed to combat multidrug-resistant bacterial infections. Application of the predatory bacterium, Bdellovibrio bacteriovorus, which kills other bacteria including pathogens, is considered promising for bacterial infections. The B. bacteriovorus life cycle consists of two phases, a free-living, invasive attack phase and an intracellular reproductive phase, in which this predatory bacterium degrades the hosts macromolecules and reuses them for its own growth. To understand the use of B. bacteriovorus as a living antibiotic, it is first necessary to dissect its life cycle including chromosome replication. Here, we present for the first time a real-time investigation into subcellular localization of chromosome replication in a single cells of B. bacteriovorus. This process initiates at the invasion pole of B. bacteriovorus and proceeds until several copies of the chromosome have been completely synthesized. Interestingly, we demonstrate that some cells of B. bacteriovorus require two prey cells sequentially to complete their life cycle.