金黄色葡萄球菌Al-2群体感应以icaR依赖的方式减弱生物膜形成
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摘要
金黄色葡萄球菌(金葡菌)是引起人类生物膜(Biofilm)感染的重要病原体。自诱导物2(Autoinducer2, AI-2)是一种负责细菌种间交流的信号分子,在革兰氏阳性和革兰氏阴性菌的群体感应(Quorum-sensing, QS)中均起着重要的调控作用,但是它在金葡菌生物膜形成中所扮演的精确角色还不清楚。本文证明了在静止、流动、厌氧和体内小鼠模型中,金葡菌RN6390B中AI-2合成酶基因luxS缺失导致生物膜形成增强,通过在luxS缺失株(ΔluxS)中外源添加化学合成的AI-2前体,可以恢复其成为野生型的生物膜表型。实时定量反转录PCR分析表明,AI-2激活了ica操纵子的一个抑制子icaR的转录,导致icaA的转录水平下降,这有可能是金葡菌中luxS缺失影响生物膜形成的主要原因。另外,我们利用AluxS,RN6911和Δagr AluxS菌株,比较了agr介导的QS系统和LuxS/AI-2QS系统在金葡菌中调节生物膜形成的关系。数据显示,agr介导的QS系统和LuxS/AI-2QS系统在金葡菌生物膜形成过程中的调控作用呈现累加效应。这些发现证明,AI-2通过一个激活icaR的途径降低金葡菌生物膜的形成。这些研究结果为金葡菌生物膜相关感染的治疗提供了线索。
Staphylococcus aureus is an important pathogen that causes biofilm-associated infection in humans. Autoinducer2(AI-2), a quorum-sensing (QS) signal for interspecies communication, has a wide range of regulatory functions in both Gram-positive and Gram-negative bacteria, but its exact role in biofilm formation in S. aureus remains unclear. Here we demonstrate that mutation of the AI-2synthase gene luxS in S. aureus RN6390B results in increased biofilm formation compared with the wild-type (WT) strain under static, flowing and anaerobic conditions and in a mouse model. Addition of the chemically synthesized AI-2precursor in the luxS mutation strain (AluxS) restored the WT phenotype. Real-time RT-PCR analysis showed that AI-2activated the transcription of icaR, a repressor of the ica operon, and subsequently a decreased level of icaA transcription, which was presumably the main reason why luxS mutation influences biofilm formation. Furthermore, we compared the roles of the agr-mediated QS system and the LuxS/AI-2QS system in the regulation of biofilm formation using the ΔluxS strain, RN6911and the Aagr AluxS strain. Our data indicate a cumulative effect of the two QS systems on the regulation of biofilm formation in S. aureus. These findings demonstrate that AI-2can decrease biofilm formation in S. aureus via an icaR-activation pathway. This study may provide clues for therapy in S. aureus biofilm-associated infection.
Staphylococcus aureus is an important pathogen that causes biofilm-associated infection in humans. Autoinducer2(AI-2), a quorum-sensing (QS) signal for interspecies communication, has a wide range of regulatory functions in both Gram-positive and Gram-negative bacteria, but its exact role in biofilm formation in S. aureus remains unclear. Here we demonstrate that mutation of the AI-2synthase gene luxS in S. aureus RN6390B results in increased biofilm formation compared with the wild-type (WT) strain under static, flowing and anaerobic conditions and in a mouse model. Addition of the chemically synthesized AI-2precursor in the luxS mutation strain (AluxS) restored the WT phenotype. Real-time RT-PCR analysis showed that AI-2activated the transcription of icaR, a repressor of the ica operon, and subsequently a decreased level of icaA transcription, which was presumably the main reason why luxS mutation influences biofilm formation. Furthermore, we compared the roles of the agr-mediated QS system and the LuxS/AI-2QS system in the regulation of biofilm formation using the ΔluxS strain, RN6911and the Aagr AluxS strain. Our data indicate a cumulative effect of the two QS systems on the regulation of biofilm formation in S. aureus. These findings demonstrate that AI-2can decrease biofilm formation in S. aureus via an icaR-activation pathway. This study may provide clues for therapy in S. aureus biofilm-associated infection.
引文
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