Wu S S, Wu J, Kaiser D
Department of Biochemistry, Stanford University School of Medicine, California 94305, USA.
Mol Microbiol. 1997 Jan;23(1):109-21. doi: 10.1046/j.1365-2958.1997.1791550.x.
Social gliding motility in Myxococcus xanthus depends on the presence of Type IV pili. To begin to examine the role of pili in social motility, 17 mutants were identified which had lost social motility, but still expressed pili. Four of these mutants carry point mutations which mapped to a locus upstream of the recently identified pilS, pilR, and pilA genes. Sequencing of this locus revealed a gene with homology to pilT from Pseudomonas Aeruginosa. Sequencing of the four point mutations revealed that they occurred within the M. xanthus pilT locus. A markerless deletion within M. xanthus pilT, similar to the four point mutations, disrupted social gliding behaviour but did not interfere with pilus formation or pilus-dependent cell-cell agglutination. Using time-lapse videomicroscopy, residual social motility was observed in dsp- strains (known to be deficient in fibril but not pilus production); this was not observed in a delta pilT dep- double mutant. Two genes flanking pilT were also sequenced, and found to have homology to pilB and pilC from P. aeruginosa. Markerless deletions within these genes caused both pilus and social-motility defects. These results indicate that M. xanthus pilB and pilC are required for pilus biogenesis, while pilT is required for assembled pili to play their role in social motility. Thus, pilB, pilT, pilC, pilS, pilR and pilA form a contiguous cluster of pil genes required for social motility.
黄色粘球菌的群体滑行运动依赖于IV型菌毛的存在。为了开始研究菌毛在群体运动中的作用,我们鉴定出了17个失去群体运动能力但仍表达菌毛的突变体。其中四个突变体携带点突变,这些突变定位在最近鉴定出的pilS、pilR和pilA基因上游的一个位点。对该位点进行测序后发现了一个与铜绿假单胞菌的pilT具有同源性的基因。对这四个点突变进行测序发现,它们发生在黄色粘球菌的pilT位点内。与这四个点突变类似,黄色粘球菌pilT内的一个无标记缺失破坏了群体滑行行为,但不影响菌毛形成或菌毛依赖性细胞间凝集。使用延时视频显微镜观察到,在dsp-菌株(已知在纤丝而非菌毛产生方面存在缺陷)中存在残余的群体运动能力;而在pilT缺失的dep-双突变体中未观察到这种情况。对pilT两侧的两个基因也进行了测序,发现它们与铜绿假单胞菌的pilB和pilC具有同源性。这些基因内的无标记缺失导致了菌毛和群体运动缺陷。这些结果表明,黄色粘球菌的pilB和pilC是菌毛生物合成所必需的,而pilT是组装好的菌毛在群体运动中发挥作用所必需的。因此,pilB、pilT、pilC、pilS、pilR和pilA形成了一个连续的菌毛基因簇,是群体运动所必需的。