Abstract
Brucella abortus is a pathogenic bacterium able to proliferate inside host cells. During the first steps of its trafficking, it is able to block the progression of its cell cycle, remaining at the G1 stage for several hours, before it reaches its replication niche. We hypothesized that starvation mediated by guanosine tetra- or penta-phosphate, (p)ppGpp, could be involved in the cell cycle arrest. Rsh is the (p)ppGpp synthetase/hydrolase. A B. abortus ∆rsh mutant is unable to grow in minimal medium, it is unable to survive in stationary phase in rich medium and it is unable to proliferate inside RAW 264.7 macrophages. A strain producing the heterologous constitutive (p)ppGpp hydrolase Mesh1b is also unable to proliferate inside these macrophages. Altogether, these data suggest that (p)ppGpp is necessary to allow B. abortus to adapt to its intracellular growth conditions. The deletion of dksA, proposed to mediate a part of the effect of (p)ppGpp on transcription, does not affect B. abortus growth in culture or inside macrophages. Expression of a gene coding for a constitutively active (p)ppGpp synthetase slows down growth in rich medium and inside macrophages. Using an mCherry–ParB fusion able to bind to the replication origin of the main chromosome of B. abortus, we observed that expression of the constitutive (p)ppGpp synthetase gene generates an accumulation of bacteria at the G1 phase. We thus propose that (p)ppGpp accumulation could be one of the factors contributing to the G1 arrest observed for B. abortus in RAW 264.7 macrophages.
Highlights
Bacteria from the Brucella genus are the causative agents of brucellosis, a neglected disease which constitutes a worldwide anthropozoonosis
Brucella abortus able to control itsreplication cell cycle progression it is inside host cells, involved in this control are unknown
Since convincing data show that and the ability adapt to starvation is a key factor for the success of cellular infections by Brucella melitensis and Brucella suis
Summary
Bacteria from the Brucella genus are the causative agents of brucellosis, a neglected disease which constitutes a worldwide anthropozoonosis. The disease is characterized by an undulant fever, named Malta fever, and in the long term the infection leads to chronicity and symptoms such as arthritis, endocarditis and can have a fatal outcome without treatment [1] In their hosts, Brucellae invade, survive and replicate inside professional and non-professional phagocytic cells such as macrophages and trophoblasts. We observed that a B. abortus dksA null mutant was able to proliferate inside host cells as the wild type (WT) strain, suggesting that DksA is not crucially involved in the (p)ppGpp-dependent phenotypes observed during infection. These results suggest that adjustment of (p)ppGpp levels are crucial for the infection process in B. abortus
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