Glucose starvation-induced dispersal of Pseudomonas aeruginosa biofilms is cAMP and energy dependent.
Glucose starvation-induced dispersal of Pseudomonas aeruginosa biofilms is cAMP and energy dependent.
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DOI：
10.1371/journal.pone.0042874
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Schleheck D
中科院分区：
文献类型：
--
作者：
Huynh TT;McDougald D;Klebensberger J;Al Qarni B;Barraud N;Rice SA;Kjelleberg S;Schleheck D
碳饥饿已被证明会诱导机会性病原体铜绿假单胞菌生物膜中的大规模扩散事件；然而，控制这种分散反应的分子途径仍然未知。我们通过差异肽指纹质谱 (iTRAQ) 量化了葡萄糖饥饿期间铜绿假单胞菌 PAO1 生物膜和浮游细胞的蛋白质组变化。此外，我们通过光度法监测分散情况，即在连续流通池中预生长和饥饿的生物膜的浊度/不透明度降低，以评估治疗方法（例如抑制剂 CCCP、砷酸盐、氯霉素、L-丝氨酸异羟肟酸盐）和生物膜发育和分散中改变的关键突变体（例如 nirS、vfr、bdlA、rpoS、 lasRrhlR、Pf4-噬菌体和 cyaA）。在野生型生物膜中，葡萄糖饥饿后 5 分钟内开始分散，2 小时后达到最大，饥饿 24 小时后高达 60% 的原始生物量已分散。蛋白质合成的变化一般不超过两倍，表明100多种属于不同类别的蛋白质存在差异表达，包括碳和能量代谢、应激适应和运动性。对于不同的处理，只有质子离子载体 CCCP 或砷酸盐（ATP 合成的抑制剂）阻止生物膜的分散。对于测试的不同突变体，只有cyaA（细胞内第二信使cAMP的合酶）未能分散； cyaA 突变的互补恢复了野生型表型。因此，铜绿假单胞菌 PAO1 中碳饥饿诱导的生物膜分散途径涉及通过直接 ATP 合成和质子动力依赖步骤产生 ATP，并通过 cAMP 介导，这可能控制参与重塑生物膜细胞以准备浮游生存的蛋白质的活性。
Carbon starvation has been shown to induce a massive dispersal event in biofilms of the opportunistic pathogen Pseudomonas aeruginosa; however, the molecular pathways controlling this dispersal response remain unknown. We quantified changes in the proteome of P. aeruginosa PAO1 biofilm and planktonic cells during glucose starvation by differential peptide-fingerprint mass-spectrometry (iTRAQ). In addition, we monitored dispersal photometrically, as a decrease in turbidity/opacity of biofilms pre-grown and starved in continuous flow-cells, in order to evaluate treatments (e.g. inhibitors CCCP, arsenate, chloramphenicol, L-serine hydroxamate) and key mutants altered in biofilm development and dispersal (e.g. nirS, vfr, bdlA, rpoS, lasRrhlR, Pf4-bacteriophage and cyaA). In wild-type biofilms, dispersal started within five minutes of glucose starvation, was maximal after 2 h, and up to 60% of the original biomass had dispersed after 24 h of starvation. The changes in protein synthesis were generally not more than two fold and indicated that more than 100 proteins belonging to various classes, including carbon and energy metabolism, stress adaptation, and motility, were differentially expressed. For the different treatments, only the proton-ionophore CCCP or arsenate, an inhibitor of ATP synthesis, prevented dispersal of the biofilms. For the different mutants tested, only cyaA, the synthase of the intracellular second messenger cAMP, failed to disperse; complementation of the cyaA mutation restored the wild-type phenotype. Hence, the pathway for carbon starvation-induced biofilm dispersal in P. aeruginosa PAO1 involves ATP production via direct ATP synthesis and proton-motive force dependent step(s) and is mediated through cAMP, which is likely to control the activity of proteins involved in remodeling biofilm cells in preparation for planktonic survival.
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影响因子：
3.2
作者：
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通讯作者：
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影响因子：
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影响因子：
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