Nelson A L, Ries J, Bagnoli F, Dahlberg S, Fälker S, Rounioja S, Tschöp J, Morfeldt E, Ferlenghi I, Hilleringmann M, Holden D W, Rappuoli R, Normark S, Barocchi M A, Henriques-Normark B
Swedish Institute for Infectious Disease Control and Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.
Mol Microbiol. 2007 Oct;66(2):329-40. doi: 10.1111/j.1365-2958.2007.05908.x. Epub 2007 Sep 10.
Adherence to host cells is important in microbial colonization of a mucosal surface, and Streptococcus pneumoniae adherence was significantly enhanced by expression of an extracellular pilus composed of three subunits, RrgA, RrgB and RrgC. We sought to determine which subunit(s) confers adherence. Bacteria deficient in RrgA are significantly less adherent than wild-type organisms, while overexpression of RrgA enhances adherence. Recombinant monomeric RrgA binds to respiratory cells, as does RrgC with less affinity, and pre-incubation of epithelial cells with RrgA reduces adherence of wild-type piliated pneumococci. Non-adherent RrgA-negative, RrgB- and RrgC-positive organisms produce pili, suggesting that pilus-mediated adherence is due to expression of RrgA, rather than the pilus backbone itself. In contrast, RrgA-positive strains with disrupted rrgB and rrgC genes exhibit wild-type adherence despite failure to produce pili by Western blot or immunoelectron microscopy. The density of bacteria colonizing the upper respiratory tract of mice inoculated with piliated RrgA-negative pneumococci was significantly less compared with wild-type; in contrast, non-piliated pneumococci expressing non-polymeric RrgA had similar numbers of bacteria in the nasopharynx as piliated wild-type bacteria. These data suggest that RrgA is central in pilus-mediated adherence and disease, even in the absence of polymeric pilus production.
对宿主细胞的黏附在微生物在黏膜表面的定殖中很重要,由三个亚基RrgA、RrgB和RrgC组成的细胞外菌毛的表达显著增强了肺炎链球菌的黏附。我们试图确定哪个亚基赋予黏附能力。缺乏RrgA的细菌比野生型生物体的黏附能力明显更低,而RrgA的过表达增强了黏附。重组单体RrgA与呼吸道细胞结合,RrgC也能结合,但亲和力较低,用RrgA预孵育上皮细胞会降低野生型有菌毛肺炎球菌的黏附。不黏附的RrgA阴性、RrgB和RrgC阳性生物体产生菌毛,这表明菌毛介导的黏附是由于RrgA的表达,而不是菌毛主干本身。相比之下,rrgB和rrgC基因被破坏的RrgA阳性菌株尽管通过蛋白质印迹或免疫电子显微镜未能产生菌毛,但仍表现出野生型黏附能力。与野生型相比,接种有菌毛RrgA阴性肺炎球菌的小鼠上呼吸道中定殖的细菌密度显著更低;相比之下,表达非聚合RrgA的无菌毛肺炎球菌在鼻咽部的细菌数量与有菌毛野生型细菌相似。这些数据表明,即使在不产生聚合菌毛的情况下,RrgA在菌毛介导的黏附及疾病中也起着核心作用。