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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

机译:布鲁氏菌流产毒力必不可少的两组分系统BvrR / BvrS调节根瘤菌科成员中外膜蛋白的表达

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摘要

The Brucella BvrR/BvrS two-component regulatory system is homologous to the ChvI/ChvG systems of Sinorhizobium meliloti and Agrobacterium tumefaciens necessary for endosymbiosis and pathogenicity in plants. BvrR/BvrS controls cell invasion and intracellular survival. Probing the surface of bvrR and bvrS transposon mutants with monoclonal antibodies showed all described major outer membrane proteins (Omps) but Omp25, a protein known to be involved in Brucella virulence. Absence of Omp25 expression was confirmed by two-dimensional electrophoresis of envelope fractions and by gene reporter studies. The electrophoretic analysis also revealed reduction or absence in the mutants of a second set of protein spots that by matrix-assisted laser desorption ionization MS and peptide mass mapping were identified as a non-previously described Omp (Omp3b). Because bvrR and bvrS mutants are also altered in cell-surface hydrophobicity, permeability, and sensitivity to surf ace-targeted bactericidal peptides, it is proposed that BvrR/BvrS controls cell envelope changes necessary to transit between extracellular and intracellular environments. A genomic search revealed that Omp25 (Omp3a) and Omp3b belong to a family of Omps of plant and animal cell-associated alpha-Proteobacteria, which includes Rhizobium leguminosarum RopB and A. tumefaciens AopB. Previous work has shown that RopB is not expressed in bacteroids, that AopB is involved in tumorigenesis, and that dysfunction of A. tumetaciens ChvI/ChvG alters surface properties. It is thus proposed that the BvrR/BvrS and Omp3 homologues of the cell-associated a-Proteobacteria play a role in bacterial surface control and host cell interactions.
机译:布鲁氏菌BvrR / BvrS两组分调节系统与植物内共生和致病性所必需的苜蓿中华根瘤菌和根癌农杆菌的ChvI / ChvG系统同源。 BvrR / BvrS控制细胞侵袭和细胞内存活。用单克隆抗体探测bvrR和bvrS转座子突变体的表面显示了所有描述的主要外膜蛋白(Omps),但Omp25是已知与布鲁氏菌毒力有关的蛋白。通过包膜部分的二维电泳和基因报告子研究证实了Omp25表达的缺失。电泳分析还显示,通过基质辅助激光解吸电离质谱和肽质量图谱鉴定为第二种蛋白斑点的突变体减少或缺失,这些蛋白斑点被确定为先前未描述的Omp(Omp3b)。由于bvrR和bvrS突变体的细胞表面疏水性,通透性和对表面靶向杀菌肽的敏感性也发生了变化,因此提出了BvrR / BvrS控制细胞外膜变化以在细胞外和细胞内环境之间迁移。基因组搜索显示,Omp25(Omp3a)和Omp3b属于与植物和动物细胞相关的α-变形杆菌属Omps家族,其中包括豆科根瘤菌RopB和根癌农杆菌AopB。先前的研究表明,RopB在类细菌中不表达,AopB与肿瘤发生有关,而根癌农杆菌ChvI / ChvG的功能障碍会改变其表面性质。因此,提出了与细胞相关的α-变形杆菌的BvrR / BvrS和Omp3同源物在细菌表面控制和宿主细胞相互作用中起作用。

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