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Backgound    Brucella melitensis  is a facultative, intracellular, pathogenic bacterium that replicates within macrophages. The type IV secretion system encoded by the  virB  operon ( virB ) is involved in  Brucella  intracellular survival. However, the underlying molecular mechanisms, especially the target proteins affected by the  virB , remain largely unclear.    Methodology/Principal Findings  In order to define the proteins affected by  virB , the proteomes of wild-type and the  virB  mutant were compared under in vitro conditions where  virB  was highly activated. The differentially expressed proteins were identified by MALDI-TOF-MS. Forty-four down-regulated and eighteen up-regulated proteins which exhibited a 2-fold or greater change were identified. These proteins included those involved in amino acid transport and metabolism, lipid metabolism, energy production, cell membrane biogenesis, translation, post-translational modifications and protein turnover, as well as unknown proteins. Interestingly, several important virulence related proteins involved in intracellular survival, including VjbR, DnaK, HtrA, Omp25, and GntR, were down-regulated in the  virB  mutant. Transcription analysis of virB and vjbR at different growth phase showed that virB positively affect transcription of vjbR in a growth phase dependent manner. Quantitative RT-PCR showed that transcription of these genes was also affected by  virB  during macrophage cell infection, consistent with the observed decreased survival of the  virB  mutant in macrophage.    Conclusions/Significance  These data indicated that the  virB  operon may control the intracellular survival of  Brucella  by affecting the expression of relevant proteins.

Comparative Proteomics Analyses Reveal the virB of B. melitensis Affects Expression of Intracellular Survival Related Proteins