Alvarez-Martinez M T, Machold J, Weise C, Schmidt-Eisenlohr H, Baron C, Rouot B
INSERM U431, Université de Montpellier 2, 34095 Montpellier Cedex 05, France.
J Bacteriol. 2001 Sep;183(18):5343-51. doi: 10.1128/JB.183.18.5343-5351.2001.
Brucella strains possess an operon encoding type IV secretion machinery very similar to that coded by the Agrobacterium tumefaciens virB operon. Here we describe cloning of the Brucella suis homologue of the chvE-gguA-gguB operon of A. tumefaciens and characterize the sugar binding protein ChvE (78% identity), which in A. tumefaciens is involved in virulence gene expression. B. suis chvE is upstream of the putative sugar transporter-encoding genes gguA and gguB, also present in A. tumefaciens, but not adjacent to that of a LysR-type transcription regulator. Although results of Southern hybridization experiments suggested that the gene is present in all Brucella strains, the ChvE protein was detected only in B. suis and Brucella canis with A. tumefaciens ChvE-specific antisera, suggesting that chvE genes are differently expressed in different Brucella species. Analysis of cell growth of B. suis and of its chvE or gguA mutants in different media revealed that ChvE exhibited a sugar specificity similar to that of its A. tumefaciens homologue and that both ChvE and GguA were necessary for utilization of these sugars. Murine or human macrophage infections with B. suis chvE and gguA mutants resulted in multiplication similar to that of the wild-type strain, suggesting that virB expression was unaffected. These data indicate that the ChvE and GguA homologous proteins of B. suis are essential for the utilization of certain sugars but are not necessary for survival and replication inside macrophages.
布鲁氏菌菌株拥有一个编码IV型分泌机制的操纵子,该操纵子与根癌农杆菌virB操纵子编码的机制非常相似。在此,我们描述了根癌农杆菌chvE-gguA-gguB操纵子的猪布鲁氏菌同源物的克隆,并对糖结合蛋白ChvE(同一性为78%)进行了表征,在根癌农杆菌中,该蛋白参与毒力基因表达。猪布鲁氏菌chvE位于假定的糖转运蛋白编码基因gguA和gguB的上游,这两个基因也存在于根癌农杆菌中,但不与LysR型转录调节因子的基因相邻。尽管Southern杂交实验结果表明该基因存在于所有布鲁氏菌菌株中,但仅在猪布鲁氏菌和犬布鲁氏菌中用根癌农杆菌ChvE特异性抗血清检测到了ChvE蛋白,这表明chvE基因在不同布鲁氏菌物种中的表达存在差异。对猪布鲁氏菌及其chvE或gguA突变体在不同培养基中的细胞生长分析表明,ChvE表现出与其根癌农杆菌同源物相似的糖特异性,并且ChvE和GguA对于这些糖的利用都是必需的。用猪布鲁氏菌chvE和gguA突变体感染小鼠或人巨噬细胞后,其增殖情况与野生型菌株相似,这表明virB表达未受影响。这些数据表明,猪布鲁氏菌的ChvE和GguA同源蛋白对于某些糖的利用至关重要,但对于在巨噬细胞内存活和复制并非必需。