近日,,國際微生物權(quán)威雜志BMC Microbiology在線刊登了美國俄勒岡州立大學(xué)研究人員的最新研究成果“Quorum sensing modulates colony morphology through alkyl quinolones in Pseudomonas aeruginosa”,文章中,,研究者揭示了綠膿桿菌的群體感應(yīng)系統(tǒng)(Quorum sensing)可以通過烷基-喹諾酮信號分子(alkyl quinolones,,AQ)來調(diào)節(jié)菌體的形態(tài)。
銅綠假單胞菌是一類重要的人類機會致病菌,,這種細菌可以寄居到多種的宿主身上,,在土壤到免疫力低下的病人身上都有分布。在環(huán)境中,,綠膿桿菌更傾向于生成生物被膜來進行繁殖生存,,并且引起感染等,細菌可以依據(jù)所處的不同環(huán)境形成不同類型的生物被膜,;在靜態(tài)的液體培養(yǎng)基表面,,細菌可以在液體-空氣交界面形成薄膜(pellicles),在流動的液體中,,細菌更傾向于吸附至固體表面,,并且在固體表面形成生物被膜,如果在完全固體的培養(yǎng)基表面,,細菌更易于生成完整形態(tài)的菌體(colonies),。生物被膜是由許多的胞外表多糖(EPS)基質(zhì)組裝而成的,而且包含了許多胞外DNA,、蛋白質(zhì),、RNA和離子等,在綠膿桿菌中,,兩個操縱子家族基因pel和psl主要合成表多糖,,前者主要編碼葡萄糖,而后者主要編碼半乳糖和甘露糖,。
綠膿桿菌的群體感應(yīng)系統(tǒng)是一個依賴細胞密度來調(diào)節(jié)的細菌信號轉(zhuǎn)導(dǎo)系統(tǒng),,細菌含有群體感應(yīng)系統(tǒng)含有三個組分:las系統(tǒng)、rhl系統(tǒng)和PQS系統(tǒng),,其中PQS系統(tǒng)以烷基-喹諾酮信號分子(AQ)為基礎(chǔ),;三個系統(tǒng)中,las系統(tǒng)(包括轉(zhuǎn)錄調(diào)節(jié)子LasR和信號合成酶LasI)中轉(zhuǎn)錄調(diào)節(jié)子LasR可以正調(diào)節(jié)AQ的生成,,而AQ同時在las系統(tǒng)缺失的情況下也可以被生成,;rhl系統(tǒng)可以抑制AQ的生成,AQ的生物合成酶可以使細菌產(chǎn)生超過50種AQ分子,,群體感應(yīng)系統(tǒng)中三個系統(tǒng)可以互相調(diào)節(jié),,他們也可以調(diào)控銅綠假單胞菌基因組超過5%的基因。
研究者Schuster Martin表示,,他們的研究工作揭示了QS系統(tǒng)和EPS之間的關(guān)系,,作者運用了染色質(zhì)免疫共沉淀微陣列法(CHIP-chip)和電泳遷移位移實驗(EMSA)等方法進行了研究,,結(jié)果表明,LasR可以結(jié)合到表多糖合成基因Psl的啟動子位置,,然而LasR的缺失如何影響EPS的產(chǎn)量以及影響菌落生物被膜的產(chǎn)生,,研究者構(gòu)建了lasR的突變體,該突變體在37℃可以形成褶皺的菌落形態(tài),,于是就表明las系統(tǒng)和psl之間有某種聯(lián)系,然后研究者發(fā)現(xiàn)菌體出現(xiàn)褶皺的形態(tài)和pel基因有關(guān)系,,而和psl基因并沒有關(guān)系,,抑制誘變實驗表明,lasR突變體涉及介導(dǎo)了PQS途徑,,研究者通過薄層色譜法對AQ分子進行了表型分析和定量分析,,結(jié)果表明AQ信號分子可以調(diào)節(jié)細菌的菌落形態(tài)。作者的研究結(jié)果表明las系統(tǒng)可以抑制Pel并且通過4-羥烷基-喹諾酮信號分子來調(diào)節(jié)細菌菌體的形態(tài),,這就揭示了銅綠假單胞菌中AQ信號分子的一個新的功能,。(生物谷:T.Shen編譯)
doi:10.1186/1471-2180-12-30
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Quorum sensing modulates colony morphology through alkyl quinolones in Pseudomonas aeruginosa
Rashmi Gupta and Martin Schuster
Background Acyl-homoserine lactone (acyl-HSL) and alkyl quinolone (AQ) based quorum-sensing (QS) systems are important for Pseudomonas aeruginosa virulence and biofilm formation. The effect of QS on biofilm formation is influenced by various genetic and environmental factors. Here, we used a colony biofilm assay to study the effect of the central acyl-HSL QS regulator, LasR, on biofilm formation and structure in the representative clinical P. aeruginosa isolate ZK2870.
Results A lasR mutant exhibited wrinkled colony morphology at 37degreesC in contrast to the smooth colony morphology of the wild-type. Mutational analysis indicated that wrinkling of the lasR mutant is dependent on pel, encoding a biofilm matrix exopolysaccharide. Suppressor mutagenesis and complementation analysis implicated the AQ signaling pathway as the link between las QS and colony morphology. In this pathway, genes pqsA-D are involved in the synthesis of 4-hydroxyalkyl quinolines ("Series A congeners"), which are converted to 3,4-dihydroxyalkyl quinolines ("Series B congeners", including the well-characterized Pseudomonas Quinolone Signal, PQS) by the product of the LasR-dependent pqsH gene. Measurement of AQ in the wild-type, the lasR pqsA::Tn suppressor mutant as well as the defined lasR, pqsH, and lasR pqsH mutants showed a correlation between 4-hydroxyalkyl quinoline levels and the degree of colony wrinkling. Most importantly, the lasR pqsH double mutant displayed wrinkly morphology without producing any 3,4-dihydroxyalkyl quinolines. Constitutive expression of pqsA-D genes in a lasR pqsR::Tn mutant showed that colony wrinkling does not require the AQ receptor PqsR.
Conclusions Taken together, these results indicate that the las QS system represses Pel and modulates colony morphology through a 4-hydroxyalkyl quinoline in a PqsR-independent manner, ascribing a novel function to an AQ other than PQS in P. aeruginosa.