Abstract
Background: In many bacteria, the phosphotransferase system (PTS) is a key player in the regulation of the assimilation of alternative carbon sources notably through catabolic repression. The intracellular pathogens Brucella spp. possess four PTS proteins (EI , NPr, EIIA and an EIIA of the mannose family) but no PTS permease suggesting that this PTS might serve only regulatory functions. Methodology/Principal Findings: In vitro biochemical analyses and in vivo detection of two forms of EIIA (phosphorylated or not) established that the four PTS proteins of Brucella melitensis form a functional phosphorelay. Moreover, in vitro the protein kinase HprK/P phosphorylates NPr on a conserved serine residue, providing an additional level of regulation to the B. melitensis PTS. This kinase activity was inhibited by inorganic phosphate and stimulated by fructose-1,6 bisphosphate. The genes encoding HprK/P, an EIIA -like protein and NPr are clustered in a locus conserved among a-proteobacteria and also contain the genes for the crucial two-component system BvrR-BvrS. RT-PCR revealed a transcriptional link between these genes suggesting an interaction between PTS and BvrR-BvrS. Mutations leading to the inactivation of EI or NPr significantly lowered the synthesis of VirB proteins, which form a type IV secretion system. These two mutants also exhibit a small colony phenotype on solid media. Finally, interaction partners of PTS proteins were identified using a yeast two hybrid screen against the whole B. melitensis ORFeome. Both NPr and HprK/P were shown to interact with an inorganic pyrophosphatase and the EIIA -like protein with the E1 component (SucA) of 2-oxoglutarate dehydrogenase. Conclusions/Significance: The B. melitensis can transfer the phosphoryl group from PEP to the EIIAs and a link between the PTS and the virulence of this organism could be established. Based on the protein interaction data a preliminary model is proposed in which this regulatory PTS coordinates also C and N metabolism.
Original language | English |
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Pages (from-to) | 1-16 |
Number of pages | 16 |
Journal | PLoS ONE |
Volume | 5 |
Issue number | 9 |
DOIs | |
Publication status | Published - 1 Jan 2010 |
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Biological Security Laboratory Level 3 (BL3)
De Bolle, X. (Manager)
Technological Platform: Biological Security Laboratory Level 3Facility/equipment: Technological Platform