Dupuy B, Pugsley A P
Unité des Neisseria, Centre National de la Recherche Scientifique URA 1149, Institut Pasteur, Paris, France.
J Bacteriol. 1994 Mar;176(5):1323-31. doi: 10.1128/jb.176.5.1323-1331.1994.
The assembly of type IV pili in Neisseria gonorrhoeae is a complex process likely to require the products of many genes. One of these is the enzyme prepilin peptidase, which cleaves and then N methylates the precursor pilin subunits prior to their assembly into pili. We have used a PCR amplification strategy to clone the N. gonorrhoeae prepilin peptidase gene, pilDNg. A single copy of the gene is shown to be present in the chromosome. Its product promotes correct cleavage of the gonococcal prepillin in Escherichia coli cells carrying both the prepilin peptidase gene and the pilin structural gene. PilDNg also cleaves prePulG, a type IV pilin-like protein of Klebsiella oxytoca. Moreover, PilDNg complements a mutation in the gene coding for the prepilin peptidase-like protein of K. oxytoca, pulO, partially restoring PulG-PulO-dependent extracellular secretion of the enzyme pullulanase. Finally, we show that genes homologous to pilDNg are present and expressed in a variety of species in the genus Neisseria, including some commensal strains.
淋病奈瑟菌IV型菌毛的组装是一个复杂的过程,可能需要许多基因的产物参与。其中之一是前菌毛蛋白肽酶,它在菌毛蛋白亚基组装成菌毛之前,先对其进行切割,然后进行N甲基化。我们采用了PCR扩增策略来克隆淋病奈瑟菌的前菌毛蛋白肽酶基因pilDNg。结果显示该基因在染色体上只有一个拷贝。它的产物能促进携带前菌毛蛋白肽酶基因和菌毛蛋白结构基因的大肠杆菌细胞中淋病奈瑟菌前菌毛蛋白的正确切割。PilDNg还能切割产酸克雷伯菌的IV型菌毛样蛋白prePulG。此外,PilDNg能弥补产酸克雷伯菌中编码前菌毛蛋白肽酶样蛋白的基因pulO的突变,部分恢复PulG - PulO依赖的淀粉酶胞外分泌。最后,我们发现与pilDNg同源的基因存在于淋病奈瑟菌属的多种物种中,包括一些共生菌株,并在这些菌株中表达。