Urban Teresa A, Goldberg Joanna B, Forstner Janet F, Sajjan Umadevi S
Department of Microbiology, University of Virginia Health Sciences, Charlottesville, Virginia, USA.
Infect Immun. 2005 Sep;73(9):5426-37. doi: 10.1128/IAI.73.9.5426-5437.2005.
Burkholderia cenocepacia strains expressing both cable (Cbl) pili and the 22-kDa adhesin bind to cytokeratin 13 (CK13) strongly and invade squamous epithelium efficiently. It has not been established, however, whether the gene encoding the adhesin is located in the cbl operon or what specific contribution the adhesin and Cbl pili lend to binding and transmigration or invasion capacity of B. cenocepacia. By immunoscreening an expression library of B. cenocepacia isolate BC7, we identified a large gene (adhA) that encodes the 22-kDa adhesin. Isogenic mutants lacking expression of either Cbl pili (cblA or cblS mutants) or the adhesin (adhA mutant) were constructed to assess the individual role of Cbl pili and the adhesin in mediating B. cenocepacia binding to and transmigration across squamous epithelium. Relative to the parent strain, mutants of Cbl pili showed reduced binding (50%) to isolated CK13, while the adhesin mutant showed almost no binding (0 to 8%). Mutants lacking either cable pili or the adhesin were compromised in their ability to bind to and transmigrate across the squamous epithelium compared to the wild-type strain, although this deficiency was most pronounced in the adhA mutant. These results indicate that both Cbl pili and the 22-kDa adhesin are necessary for the optimal binding to CK13 and transmigration properties of B. cenocepacia.
同时表达丝状(Cbl)菌毛和22 kDa黏附素的洋葱伯克霍尔德菌菌株能与细胞角蛋白13(CK13)紧密结合,并有效侵入鳞状上皮。然而,编码该黏附素的基因是否位于cbl操纵子中,以及该黏附素和Cbl菌毛对洋葱伯克霍尔德菌的结合、迁移或侵袭能力有何具体贡献,目前尚未明确。通过对洋葱伯克霍尔德菌分离株BC7的表达文库进行免疫筛选,我们鉴定出一个编码22 kDa黏附素的大基因(adhA)。构建了缺乏Cbl菌毛表达(cblA或cblS突变体)或黏附素表达(adhA突变体)的同基因突变体,以评估Cbl菌毛和黏附素在介导洋葱伯克霍尔德菌与鳞状上皮结合及跨上皮迁移中的各自作用。相对于亲本菌株,Cbl菌毛突变体与分离的CK13的结合减少(50%),而黏附素突变体几乎没有结合(0%至8%)。与野生型菌株相比,缺乏丝状菌毛或黏附素的突变体在与鳞状上皮结合和跨上皮迁移的能力上受到损害,尽管这种缺陷在adhA突变体中最为明显。这些结果表明,Cbl菌毛和22 kDa黏附素对于洋葱伯克霍尔德菌与CK13的最佳结合及迁移特性都是必需的。