National Reference Laboratory for Campylobacter, Department of Biological Safety, German Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany.
Institute of Food Safety and Food Hygiene, Freie Universität Berlin, Königsweg 69, 14163 Berlin, Germany.
Biomolecules. 2023 Mar 11;13(3):514. doi: 10.3390/biom13030514.
is an important food-borne pathogen displaying high genetic diversity, substantially based on natural transformation. The mechanism of DNA uptake from the environment depends on a type II secretion/type IV pilus system, whose components are partially known. Here, we quantified DNA uptake in at the single cell level and observed median transport capacities of approximately 30 kb per uptake location. The process appeared to be limited by the initialization of DNA uptake, was finite, and, finalized within 30 min of contact to DNA. Mutants lacking either the outer membrane pore PilQ or the inner membrane channel ComEC were deficient in natural transformation. The periplasmic DNA binding protein ComE was negligible for DNA uptake, which is in contrast to its proposed function. Intriguingly, a mutant lacking the unique periplasmic protein Cj0683 displayed rare but fully functional DNA uptake events. We conclude that Cj0683 was essential for the efficient initialization of DNA uptake, consistent with the putative function as a competence pilus protein. Unravelling features important in natural transformation might lead to target identification, reducing the adaptive potential of pathogens.
是一种重要的食源性病原体,具有很高的遗传多样性,主要基于自然转化。从环境中摄取 DNA 的机制依赖于 II 型分泌/IV 型菌毛系统,其部分成分是已知的。在这里,我们在单细胞水平上定量了 在 中的 DNA 摄取,并观察到每个摄取位置的平均转运能力约为 30kb。这个过程似乎受到 DNA 摄取初始化的限制,是有限的,并在与 DNA 接触 30 分钟内完成。缺乏外膜孔 PilQ 或内膜通道 ComEC 的突变体在自然转化中缺失。周质 DNA 结合蛋白 ComE 对 DNA 摄取的作用可以忽略不计,这与它的预期功能相反。有趣的是,一个缺乏独特的周质蛋白 Cj0683 的突变体显示出罕见但完全功能的 DNA 摄取事件。我们得出结论,Cj0683 对于 DNA 摄取的有效初始化是必不可少的,这与作为感受态菌毛蛋白的假定功能一致。揭示自然转化中重要的特征可能会导致目标的识别,从而降低病原体的适应性潜力。