Alm R A, Bodero A J, Free P D, Mattick J S
Centre for Molecular and Cellular Biology, University of Queensland, Brisbane, Australia.
J Bacteriol. 1996 Jan;178(1):46-53. doi: 10.1128/jb.178.1.46-53.1996.
The opportunistic pathogen Pseudomonas aeruginosa produces type 4 fimbriae which promote adhesion to epithelial cells and are associated with a form of surface translocation called twitching motility. We have used transposon mutagenesis to identify loci required for fimbrial assembly or function by screening for mutants that lack the spreading colony morphology characteristic of twitching motility. A subset of these mutants is resistant to fimbria-specific phage. One of these mutants (R270) was found to contain a transposon insertion in a new gene, termed pilZ, which is located on chromosomal SpeI fragment I at about 40 min on the P. aeruginosa map, a position remote from other loci involved in fimbrial biogenesis. pilZ appears to be linked to and possibly forms an operon with a gene, holB*, which is homologous to the gene encoding the delta' subunit of Escherichia coli DNA polymerase III. The product of the pilZ gene is a protein of 118 amino acids (predicted molecular weight, 12,895) which probably has a cytoplasmic location. PilZ appears to be a new class of protein which has not hitherto been represented in the sequence databases, and its function is unknown. Complementation studies indicate that pilZ is able to restore the expression of fimbriae on the surface of P. aeruginosa, as well as twitching motility and sensitivity to fimbria-specific phage when provided in trans to the R270 mutant.
机会致病菌铜绿假单胞菌产生4型菌毛,这种菌毛可促进其黏附于上皮细胞,并与一种称为“颤动运动”的表面移位形式有关。我们利用转座子诱变技术,通过筛选缺乏颤动运动所特有的扩散菌落形态特征的突变体,来鉴定菌毛组装或功能所需的基因座。这些突变体中的一部分对菌毛特异性噬菌体具有抗性。其中一个突变体(R270)被发现含有一个转座子插入到一个新基因中,该基因被命名为pilZ,它位于铜绿假单胞菌染色体SpeI片段I上,在铜绿假单胞菌图谱上约40分钟处,该位置远离其他参与菌毛生物合成的基因座。pilZ似乎与一个基因holB相连,并且可能形成一个操纵子,holB与编码大肠杆菌DNA聚合酶III的δ'亚基的基因同源。pilZ基因的产物是一种由118个氨基酸组成的蛋白质(预测分子量为12,895),可能定位于细胞质中。PilZ似乎是一类新的蛋白质,在序列数据库中尚未有相关记载,其功能未知。互补实验表明,当将pilZ基因反式提供给R270突变体时,它能够恢复铜绿假单胞菌表面菌毛的表达、颤动运动以及对菌毛特异性噬菌体的敏感性。