BvrR/BvrS-controlled outer membrane proteins Omp3a and Omp3b are not essential for Brucella abortus virulence.
The Brucella abortus two-component regulatory system BvrR/BvrS controls the expression of outer membrane proteins (Omp) Omp3a (Omp25) and Omp3b (Omp22). Disruption of bvrS or bvrR generates avirulent mutants with altered cell permeability, higher sensitivity to microbicidal peptides, and complement. Consequently, the role of Omp3a and Omp3b in virulence was examined. Similar to bvrS or bvrR mutants, omp3a and omp3b mutants displayed increased attachment to cells, indicating surface alterations. However, they showed unaltered permeability; normal expression of Omp10, Omp16, Omp19, Omp2b, and Omp1; native hapten polysaccharide; and lipopolysaccharide and were resistant to complement and polymyxin B at ranges similar to those of the wild-type (WT) counterpart. Likewise, omp3a and omp3b mutants were able to replicate in murine macrophages and in HeLa cells, were resistant to the killing action of human neutrophils, and persisted in mice, like the WT strain. Murine macrophages infected with the omp3a mutant generated slightly higher levels of tumor necrosis factor alpha than the WT, whereas the bvrS mutant induced lower levels of this cytokine. Since the absence of Omp3a or Omp3b does not result in attenuation, it can be concluded that BvrR/BvrS influences additional Brucella properties involved in virulence. Our results are discussed in the light of previous works suggesting that disruption of omp3a generates attenuated Brucella strains, and we speculate on the role of group 3 Omps.
Departamento de Microbiología y Parasitología, Universidad de Navarra, 31008 Pamplona, Spain.
Lorea
L
Manterola
Caterina
C
Guzmán-Verri
Esteban
E
Chaves-Olarte
Elías
E
Barquero-Calvo
María-Jesús
MJ
de Miguel
Ignacio
I
Moriyón
María-Jesús
MJ
Grilló
Ignacio
I
López-Goñi
Edgardo
E
Moreno
D000818:Animals; D001422:Bacterial Adhesion / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000502:physiology*; D001426:Bacterial Proteins / Q000502:physiology; D002003:Brucella abortus / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity* / Q000502:physiology; D002006:Brucellosis; D002460:Cell Line; D015169:Colony Count, Microbial; D005260:Female; D017353:Gene Deletion; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D016254:Mutagenesis, Insertional; D009504:Neutrophils / Q000276:immunology; D013154:Spleen / Q000382:microbiology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D014774:Virulence / Q000235:genetics*; D037521:Virulence Factors / Q000235:genetics / Q000502:physiology*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C471339:BvrR protein, Brucella; C471340:BvrS protein, Brucella; D014409:Tumor Necrosis Factor-alpha; D037521:Virulence Factors