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首页> 外文期刊>Journal of bacteriology >Type IV Pilus Genes pilA andpilC of Pseudomonas stutzeri Are Required for Natural Genetic Transformation, and pilA Can Be Replaced by Corresponding Genes from Nontransformable Species
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Type IV Pilus Genes pilA andpilC of Pseudomonas stutzeri Are Required for Natural Genetic Transformation, and pilA Can Be Replaced by Corresponding Genes from Nontransformable Species

机译：自然遗传转化需要斯图氏假单胞菌的IV型毛发基因pilA和pilC，并且pilA可以被非转化物种的相应基因替代

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摘要

Pseudomonas stutzeri lives in terrestrial and aquatic habitats and is capable of natural genetic transformation. After transposon mutagenesis, transformation-deficient mutants were isolated from a P. stutzeri JM300 strain. In one of them a gene which coded for a protein with 75% amino acid sequence identity to PilC of Pseudomonas aeruginosa, an accessory protein for type IV pilus biogenesis, was inactivated. The presence of type IV pili was demonstrated by susceptibility to the type IV pilus-dependent phage PO4, by occurrence of twitching motility, and by electron microscopy. The pilC mutant had no pili and was defective in twitching motility. Further sequencing revealed that pilC is clustered in an operon with genes homologous to pilB andpilD of P. aeruginosa, which are also involved in pilus formation. Next to these genes but transcribed in the opposite orientation a pilA gene encoding a protein with high amino acid sequence identity to pilin, the structural component of type IV pili, was identified. Insertional inactivation of pilAabolished pilus formation, PO4 plating, twitching motility, and natural transformation. The amounts of 3H-labeled P. stutzeri DNA that were bound to competent parental cells and taken up were strongly reduced in the pilC andpilA mutants. Remarkably, the cloned pilA genes from nontransformable organisms like Dichelobacter nodosusand the PAK and PAO strains of P. aeruginosa fully restored pilus formation and transformability of the P. stutzeri pilA mutant (along with PO4 plating and twitching motility). It is concluded that the type IV pili of the soil bacterium P. stutzeri function in DNA uptake for transformation and that their role in this process is not confined to the species-specific pilin.
机译： Pseudomonas stutzeri 生活在陆地和水生生境中，并且能够自然遗传转化。转座子诱变后，从 P中分离出转化缺陷型突变体。 stutzeri JM300菌株。其中一个基因编码的蛋白质与铜绿假单胞菌的PilC具有75％的氨基酸序列同一性，该铜绿假单胞菌是IV型菌毛生物发生的辅助蛋白，该基因被灭活。 IV型菌毛的存在通过对IV型菌毛依赖性噬菌体PO4的易感性，发生抽动运动和电子显微镜证明。 pilC 突变体没有菌毛，并且抽动力缺陷。进一步测序表明， pilC 聚集在一个操纵子中，其基因与 P的 pilB 和 pilD 同源。铜绿菌，也参与菌毛的形成。在这些基因旁边，但转录方向相反，鉴定出了一种编码 pilA 基因，该基因编码与pilin（IV型菌毛的结构成分）具有高度氨基酸序列同一性的蛋白质。 pilA 的插入失活消除了菌毛的形成，PO4电镀，抽搐运动和自然转化。 3 H标记的 P的数量。 pilC 和 pilA 突变体中，与感受态亲本细胞结合并被摄取的stutzeri DNA明显减少。值得注意的是，克隆的 pilA 基因是从不可转化的生物（如双歧杆菌）和 P的PAK和PAO菌株中克隆的。铜绿菌完全恢复了 P的菌毛形成和可转化性。 stutzeri pilA 突变体（以及PO4镀层和抽搐运动性）。结论是土壤细菌 P的IV型菌毛。 stutzeri 在转化过程中对DNA的吸收中起作用，并且它们在此过程中的作用并不局限于特定物种的菌毛蛋白。

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