Ryll R R, Rudel T, Scheuerpflug I, Barten R, Meyer T F
Max-Planck-Institut für Biologie, Abteilung Infektionsbiologie, Tübingen, Germany.
Mol Microbiol. 1997 Mar;23(5):879-92. doi: 10.1046/j.1365-2958.1997.2631630.x.
Type 4 pili produced by the pathogenic Neisseria species constitute primary determinants for the adherence to host tissues. In addition to the major pilin subunit (PilE), neisserial pili contain the variable PilC proteins represented by two variant gene copies in most pathogenic Neisseria isolates. Based upon structural differences in the conserved regions of PilE, two pilus classes can be distinguished in Neisseria meningitidis. For class I pili found in both Neisseria gonorrhoeae and N. meningitidis, PilC proteins have been implicated in pilus assembly, natural transformation competence and adherence to epithelial cells. In this study, we used primers specific for the pilC2 gene of N.gonorrhoeae strain MS11 to amplify, by the polymerase chain reaction, and clone a homologous pilC gene from N. meningitidis strain A1493 which produces class II pili. This gene was sequenced and the deduced amino acid sequence showed 75.4% and 73.8% identity with the gonococcal PilC1 and PilC2, respectively. These values match the identity value of 74.1% calculated for the two N. gonorrhoeae MS11 PilC proteins, indicating a horizontal relationship between the N. gonorrhoeae and N. meningitidis pilC genes. We provide evidence that PilC functions in meningococcal class II pilus assembly and adherence. Furthermore, expression of the cloned N. meningitidis pilC gene in a gonococcal pilC1,2 mutant restores pilus assembly, adherence to ME-180 epithelial cells, and transformation competence to the wild-type level. Thus, PilC proteins exhibit indistinguishable functions in the context of class I and class II pili.
致病性奈瑟菌属产生的4型菌毛是其黏附宿主组织的主要决定因素。除了主要菌毛亚基(PilE)外,奈瑟菌菌毛还含有可变的PilC蛋白,在大多数致病性奈瑟菌分离株中由两个变异基因拷贝代表。基于PilE保守区域的结构差异,在脑膜炎奈瑟菌中可区分出两类菌毛。对于在淋病奈瑟菌和脑膜炎奈瑟菌中均发现的I类菌毛,PilC蛋白与菌毛组装、自然转化能力以及对上皮细胞的黏附有关。在本研究中,我们使用针对淋病奈瑟菌菌株MS11的pilC2基因的特异性引物,通过聚合酶链反应进行扩增,并从产生II类菌毛的脑膜炎奈瑟菌菌株A1493中克隆出一个同源pilC基因。对该基因进行测序,推导的氨基酸序列与淋病奈瑟菌的PilC1和PilC2分别具有75.4%和73.8%的同一性。这些值与为淋病奈瑟菌MS11的两种PilC蛋白计算的74.1%的同一性值相匹配,表明淋病奈瑟菌和脑膜炎奈瑟菌的pilC基因之间存在水平关系。我们提供证据表明PilC在脑膜炎奈瑟菌II类菌毛组装和黏附中发挥作用。此外,在淋病奈瑟菌pilC1,2突变体中表达克隆的脑膜炎奈瑟菌pilC基因可将菌毛组装、对ME-180上皮细胞的黏附以及转化能力恢复到野生型水平。因此,PilC蛋白在I类和II类菌毛的背景下表现出难以区分的功能。