The type IV secretion system affects the expression of Omp25/Omp31 and the outer membrane properties of Brucella melitensis.

The type IV secretion system (T4SS) contributes to Brucella intracellular survival through its effector proteins. Comparative proteomic analysis showed that intracellular survival proteins are expressed differentially in a virB mutant. Interestingly, several outer membrane proteins (OMPs) are also differentially expressed, implying that T4SS might affect the OM properties of Brucella. To further evaluate the impact of T4SS on OM, in the present study, the OM proteomes were isolated and compared. Many more products of OMPs, particularly different products of the Omp25/Omp31 family, were found to be altered in the virB mutant. The transcription profiles of Omp25/Omp31 were different from those of their protein products, implying their regulation by virB at both transcriptional and post-transcriptional levels. The virB mutant aggregates at a high cell density and produces exopolysaccharide, a phenotype resembling that of the vjbR mutant. The virB mutant showed increased sensitivity to polymyxin B and decreased survival under oxidative, high-salt and high-osmolarity stresses, indicating drastic membrane alterations. These results indicated that in addition to being an effector protein secretion system, T4SS affects OM properties that might be important for the adaptation of Brucella to both in vitro and in vivo hostile environments.

[1]  Yufei Wang,et al.  Cytotoxicity of Brucella smooth strains for macrophages is mediated by increased secretion of the type IV secretion system. , 2009, Microbiology.

[2]  V. DelVecchio,et al.  Differential composition of culture supernatants from wild-type Brucella abortus and its isogenic virB mutants , 2009, Archives of Microbiology.

[3]  Ruifu Yang,et al.  Comparative Proteomics Analyses Reveal the virB of B. melitensis Affects Expression of Intracellular Survival Related Proteins , 2009, PloS one.

[4]  J. V. van Dijl,et al.  Identification of VceA and VceC, two members of the VjbR regulon that are translocated into macrophages by the Brucella type IV secretion system , 2008, Molecular microbiology.

[5]  A. Cloeckaert,et al.  Role of the Omp25/Omp31 Family in Outer Membrane Properties and Virulence of Brucella ovis , 2007, Infection and Immunity.

[6]  J. Lemaire,et al.  Mutations of the Quorum Sensing-Dependent Regulator VjbR Lead to Drastic Surface Modifications in Brucella melitensis , 2007, Journal of bacteriology.

[7]  Joseph P. Connolly,et al.  Proteomic analysis of Brucella abortus cell envelope and identification of immunogenic candidate proteins for vaccine development , 2006, Proteomics.

[8]  Liuyu Huang,et al.  Immunoproteomics of outer membrane proteins and extracellular proteins of Shigella flexneri 2a 2457T , 2005, Proteomics.

[9]  M. Koch,et al.  The Lipopolysaccharide of Brucella abortus BvrS/BvrR Mutants Contains Lipid A Modifications and Has Higher Affinity for Bactericidal Cationic Peptides , 2005, Journal of bacteriology.

[10]  J. Letesson,et al.  A quorum‐sensing regulator controls expression of both the type IV secretion system and the flagellar apparatus of Brucella melitensis , 2005, Cellular microbiology.

[11]  T. Ficht Intracellular survival of Brucella: defining the link with persistence. , 2003, Veterinary microbiology.

[12]  J. Paquet,et al.  Major outer membrane proteins of Brucella spp.: past, present and future. , 2002, Veterinary microbiology.

[13]  J. Garin,et al.  The two-component system BvrR/BvrS essential for Brucella abortus virulence regulates the expression of outer membrane proteins with counterparts in members of the Rhizobiaceae , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. Watarai,et al.  Modulation of Brucella‐induced macropinocytosis by lipid rafts mediates intracellular replication , 2002, Cellular microbiology.

[15]  Shouxiong Huang,et al.  Outer membrane proteins: key players for bacterial adaptation in host niches. , 2002, Microbes and infection.

[16]  D. Sánchez,et al.  A Homologue of an Operon Required for DNA Transfer in Agrobacterium Is Required in Brucella abortusfor Virulence and Intracellular Multiplication , 2000, Journal of bacteriology.

[17]  D. O’Callaghan,et al.  A homologue of the Agrobacterium tumefaciens VirB and Bordetella pertussis Ptl type IV secretion systems is essential for intracellular survival of Brucella suis , 1999, Molecular microbiology.

[18]  R. Parton,et al.  Brucella abortus Transits through the Autophagic Pathway and Replicates in the Endoplasmic Reticulum of Nonprofessional Phagocytes , 1998, Infection and Immunity.

[19]  I. Moriyón,et al.  Structure and properties of the outer membranes of Brucella abortus and Brucella melitensis. , 1998, International microbiology : the official journal of the Spanish Society for Microbiology.

[20]  A. Weintraub,et al.  Brucella-Salmonella lipopolysaccharide chimeras are less permeable to hydrophobic probes and more sensitive to cationic peptides and EDTA than are their native Brucella sp. counterparts , 1996, Journal of bacteriology.

[21]  E. Moreno,et al.  The outer membranes of Brucella spp. are resistant to bactericidal cationic peptides , 1995, Infection and immunity.

[22]  I. Moriyón,et al.  The outer membranes of Brucella spp. are not barriers to hydrophobic permeants , 1993, Journal of bacteriology.