Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.
Department of Systems Biology, Technical University of Denmark, Lyngby, Denmark.
Microbiologyopen. 2017 Dec;6(6). doi: 10.1002/mbo3.516. Epub 2017 Aug 6.
Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun-nup regulon. Organization of the regulon is similar to clusters found in other CLP-producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation was reduced in a nunF knockout mutant suggesting that these CLPs may also play a role in these phenomena as observed in other pseudomonads. Fusion of the nunF promoter region to mCherry showed that nunF is strongly upregulated in response to carbon sources indicating the presence of a fungus suggesting that environmental elicitors may also influence nunF expression which upon activation regulates nunamycin and nunapeptin production required for the growth inhibition of phytopathogens.
农霉素和农那肽是荧光假单胞菌 In5 产生的两种抗菌环脂肽(CLP),由位于两个基因簇上的非核糖体合成酶(NRPS)合成,这些基因簇被指定为 nun-nup 调控子。调控子的组织类似于其他产生 CLP 的假单胞菌中的簇,除了位于假定的 LuxR 型调节剂的边界区域。本研究重点研究了 LuxR 型编码基因 nunF 在荧光假单胞菌 In5 的 CLP 生产中的调节作用。nunF 的功能分析与液相色谱-高分辨率质谱(LC-HRMS)相结合表明,CLP 生物合成受 nunF 调节。实时定量 PCR 分析表明,当 nunF 突变时,催化 CLP 生产的 NRPS 基因的转录强烈减少,表明 nunF 是 nun-nup 调控子的一部分。在 nunF 敲除突变体中,群集和生物膜形成减少,表明这些 CLP 也可能在其他假单胞菌中观察到的这些现象中发挥作用。nunF 启动子区域与 mCherry 的融合表明,nunF 强烈上调响应碳源表明存在真菌表明环境刺激物也可能影响 nunF 表达,其激活后调节农霉素和农那肽的产生,这是抑制植物病原体生长所必需的。