Differential impact on motility and biofilm dispersal of closely related phosphodiesterases in Pseudomonas aeruginosa
Differential impact on motility and biofilm dispersal of closely related phosphodiesterases in Pseudomonas aeruginosa
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DOI：
10.1038/s41598-020-63008-5
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发表时间：
2020-04-10
影响因子：
4.6
通讯作者：
Tews, Ivo
中科院分区：
文献类型：
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作者：
Cai, Yu-ming;Hutchin, Andrew;Tews, Ivo
在铜绿假单胞菌中，浮游生活方式和生物膜生活方式之间的转变由细胞内第二信使环状二聚体-GMP (c-di-GMP) 响应环境条件进行调节。在这里，我们使用基因删除来研究环境刺激一氧化氮 (NO) 如何与生物膜分散相关，重点关注含有假定的 c-di-GMP 转换和 Per-Arnt-Sim (PAS) 感觉域的蛋白质的生物膜分散表型。我们记录了基因 Delta rbdA 和 Delta pa2072 的相反生理作用，它们编码具有相同结构域结构的蛋白质：与野生型相比，Delta rbdA 显示 c-di-GMP 水平升高、运动受限并促进生物膜形成，而 Delta pa2072 中 c-di-GMP 水平降低，生物膜形成受到抑制。根据预测的 c-di-GMP 周转功能受损，选择了第二对基因 Delta fimX 和 Delta dialA：Delta fimX 显示增加，Delta dialA 减少 NO 诱导的生物膜分散，并且这些基因影响不同类型的运动，Delta fimX 的抽搐减少，Delta dialA 的游泳减少。对于所有四种缺失突变体，我们发现NO诱导的生物量减少与NO驱动的集群增加相关，强调了这种运动在生物膜分散中的重要作用。因此，铜绿假单胞菌能够使用结构高度相关的蛋白质来区分 c-di-GMP 输出，这些蛋白质可以包含简并的 c-di-GMP 转换结构域。
In Pseudomonas aeruginosa, the transition between planktonic and biofilm lifestyles is modulated by the intracellular secondary messenger cyclic dimeric-GMP (c-di-GMP) in response to environmental conditions. Here, we used gene deletions to investigate how the environmental stimulus nitric oxide (NO) is linked to biofilm dispersal, focusing on biofilm dispersal phenotype from proteins containing putative c-di-GMP turnover and Per-Arnt-Sim (PAS) sensory domains. We document opposed physiological roles for the genes Delta rbdA and Delta pa2072 that encode proteins with identical domain structure: while Delta rbdA showed elevated c-di-GMP levels, restricted motility and promoted biofilm formation, c-di-GMP levels were decreased in Delta pa2072, and biofilm formation was inhibited, compared to wild type. A second pair of genes, Delta fimX and Delta dipA, were selected on the basis of predicted impaired c-di-GMP turnover function: Delta fimX showed increased, Delta dipA decreased NO induced biofilm dispersal, and the genes effected different types of motility, with reduced twitching for Delta fimX and reduced swimming for Delta dipA. For all four deletion mutants we find that NO-induced biomass reduction correlates with increased NO-driven swarming, underlining a significant role for this motility in biofilm dispersal. Hence P. aeruginosa is able to differentiate c-di-GMP output using structurally highly related proteins that can contain degenerate c-di-GMP turnover domains.