In addition to the Omp25/Omp31 family, iron-regulated OMP FrpB (BMEII0105), organic solvent tolerance protein Imp (BMEI1264), metal chelate OM receptor BtuB (BMEI0657) and an unknown OMP BMEI1895 were also downregulated in the virB mutant. The decreased expression of these OMPs implies that
inactivation of T4SS may lead to a drastic surface modification in B. melitensis. Biofilm is a growth form where the bacteria cells are embedded in a matrix, providing some advantages for Tofacitinib manufacturer bacteria adaptation to different environments. Brucella is also able to form a biofilm, as exemplified by a vjbR mutant. The vjbR mutant clumps at a high cell density and produces exopolysaccharides, a component of biofilm extracellular matrices. The production of exopolysaccharides was related to Omp31, as a double mutant of vjbR and omp31 was unable to produce exopolysaccharides. Dot blot showed that the vjbR-deficient strain exhibited decreased production of Omp25 and
Omp31, and increased expression of Omp10, Omp19 and Omp89, indicating that mutation of vjbR considerably modifies the membrane structure (Uzureau et al., 2007). We Navitoclax unexpectedly found that the virB mutants also form aggregates resembling those formed by the vjbR mutant, forming the aggregates at a high cell density and producing exopolysaccharides in the extracellular matrices. The membrane structure modification occurring in the virB mutant was also consistent with that of the vjbR mutant: downregulation of Omp25 and Omp31. In addition, we found that different products of the Omp25/Omp31
family were differentially expressed, and their transcription was altered, indicating that the virB affected expression of Omp25 and Omp31 at both transcriptional BCKDHB and post-translational levels. Taking into account our previous result of the positive regulation of vjbR by virB, it is possible that the aggregation, production of exopolysaccharides and decreased expression of Omp25/Omp31 might be the result of a decreased expression of vjbR in the virB mutant. However, this does not exclude other possibilities. For example, biofilm formation is a very complicated process involving a large set of genes. Membrane structure and metabolism-related genes are involved in biofilm formation. Actually, in our previous results, we found that T4SS affects the expression of many metabolism-related genes. The particular properties of the OM are thought to be responsible for the resistance of Brucella spp. to the bactericidal action of cationic peptides (Martinez de Tejada & Moriyon, 1993; Freer et al., 1996). To determine the stability of the OM, the susceptibility of the virB mutant to polymyxin B was assayed. The results showed that the virB mutant was more susceptible to polymyxin B compared with B. melitensis wild-type and the complementary strains (Fig. 4a).