Institut Pasteur, Unité de Biologie des Spirochètes, Paris, France.
Infect Immun. 2012 Jun;80(6):2019-25. doi: 10.1128/IAI.00131-12. Epub 2012 Mar 26.
Spirochetes have periplasmic flagella composed of a core surrounded by a sheath. The pathogen Leptospira interrogans has four flaB (proposed core subunit) and two flaA (proposed sheath subunit) genes. The flaA genes are organized in a locus with flaA2 immediately upstream of flaA1. In this study, flaA1 and flaA2 mutants were constructed by transposon mutagenesis. Both mutants still produced periplasmic flagella. The flaA1 mutant did not produce FlaA1 but continued to produce FlaA2 and retained normal morphology and virulence in a hamster model of infection but had reduced motility. The flaA2 mutant did not produce either the FlaA1 or the FlaA2 protein. Cells of the flaA2 mutant lacked the distinctive hook-shaped ends associated with L. interrogans and lacked translational motility in liquid and semisolid media. These observations were confirmed with a second, independent flaA2 mutant. The flaA2 mutant failed to cause disease in animal models of acute infection. Despite lacking FlaA proteins, the flagella of the flaA2 mutant were of the same thickness as wild-type flagella, as measured by electron microscopy, and exhibited a normal flagellum sheath, indicating that FlaA proteins are not essential for the synthesis of the flagellum sheath, as observed for other spirochetes. This study shows that FlaA subunits contribute to leptospiral translational motility, cellular shape, and virulence.
螺旋体具有由核心环绕的周质鞭毛,由鞘包围。病原体问号钩端螺旋体有四个 flaB(提议的核心亚基)和两个 flaA(提议的鞘亚基)基因。flaA 基因组织在一个位点中,flaA2 紧邻 flaA1 的上游。在这项研究中,通过转座子诱变构建了 flaA1 和 flaA2 突变体。这两个突变体仍产生周质鞭毛。flaA1 突变体不产生 FlaA1,但继续产生 FlaA2,并在仓鼠感染模型中保持正常形态和毒力,但运动性降低。flaA2 突变体既不产生 FlaA1 也不产生 FlaA2 蛋白。flaA2 突变体细胞缺乏与问号钩端螺旋体相关的独特钩状末端,并且在液体和半固体培养基中缺乏翻译运动性。这些观察结果通过第二个独立的 flaA2 突变体得到了证实。flaA2 突变体在急性感染动物模型中不能引起疾病。尽管缺乏 FlaA 蛋白,但 flaA2 突变体的鞭毛厚度与野生型鞭毛相同,如电子显微镜测量所示,并且表现出正常的鞭毛鞘,表明 FlaA 蛋白对于鞭毛鞘的合成不是必需的,如其他螺旋体观察到的那样。这项研究表明 FlaA 亚基有助于钩端螺旋体的翻译运动性、细胞形状和毒力。