Xu Cao, Li Man, Huang Yuanyuan, Zhang Zhe, Bian Zirui, Song Shuishan
Biology Institute, Hebei Academy of Sciences, Shijiazhuang 050051, China.
Wei Sheng Wu Xue Bao. 2011 Jun;51(6):769-75.
Quorum-sensing (QS) is a regulatory mechanism with which bacteria regulate the gene expression according to their population density. Pseudomonas aeruginosa regulates the expression of multiple genes via a hierarchical quorum-sensing cascade through LasR and RhlR and their cognate signal molecules N-(3-oxododecanoyl)-L-homoserine lactone (30-C12-HSL) and N-(butanoyl)-L-homoserine lactone (C4-HSL).
It aims to explore the regulation of QS on biosynthesis of polyhydroxyalkanoates (PHA) in P. aeruginosa.
Wild-type P. aeruginosa PA01 and its QS mutants were used to investigate the effects of quorum-sensing on biosynthesis of PHA by GC and real-time PCR at physiological and molecular level.
After treated with QS signal molecule synthesis inhibitor azithromycin, the accumulation of PHA significantly decreased in P. aeruginosa PA01 and its QS mutant strains. The content of PHA in C4-HSL synthase gene rhlI mutant strain PA0210 had no significant difference compared with that of the wild type. However, the PHA contents were significantly affected in 30C12-HSL synthase gene lasl mutant strain PA055, 30C12-HSL transcriptional regulator gene lasR mutant strain PA056 and lasI/lasR double mutant strain PA057. PHA synthase gene phaC1 expression exhibited a significant reduction in lasI mutant and lasR mutant strains. 30C12-HSL signal molecules complementary experiment shows that the expression of phaC1 can be recovered to the level of the wild type, but the synthesis of PHA is only partially restored in lasI mutant strain.
The results implicates that lasI/lasR system might be involved in the regulation of intracellular PHA biosynthesis in P. aeruginosa PA01.
群体感应(QS)是一种细菌根据其群体密度调节基因表达的调控机制。铜绿假单胞菌通过LasR和RhlR及其同源信号分子N-(3-氧代十二烷酰基)-L-高丝氨酸内酯(30-C12-HSL)和N-(丁酰基)-L-高丝氨酸内酯(C4-HSL),经由分层的群体感应级联反应调节多个基因的表达。
旨在探究群体感应对铜绿假单胞菌中聚羟基脂肪酸酯(PHA)生物合成的调控作用。
使用野生型铜绿假单胞菌PA01及其群体感应突变体,通过气相色谱法和实时定量聚合酶链反应,在生理和分子水平上研究群体感应对PHA生物合成的影响。
用群体感应信号分子合成抑制剂阿奇霉素处理后,铜绿假单胞菌PA01及其群体感应突变体菌株中PHA的积累显著减少。C4-HSL合酶基因rhlI突变体菌株PA0210中PHA的含量与野生型相比无显著差异。然而,30C12-HSL合酶基因lasl突变体菌株PA055、30C12-HSL转录调节基因lasR突变体菌株PA056和lasI/lasR双突变体菌株PA057中PHA的含量受到显著影响。PHA合酶基因phaC1的表达在lasI突变体和lasR突变体菌株中显著降低。30C12-HSL信号分子互补实验表明,phaC1的表达可恢复到野生型水平,但在lasI突变体菌株中PHA的合成仅部分恢复。
结果表明lasI/lasR系统可能参与铜绿假单胞菌PA01细胞内PHA生物合成的调控。