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鉴定和表征假交替单胞菌中的 LuxI/R 型群体感应系统。

Identification and characterization of a LuxI/R-type quorum sensing system in Pseudoalteromonas.

机构信息

College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, China.

College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, China.

出版信息

Res Microbiol. 2019 Sep-Oct;170(6-7):243-255. doi: 10.1016/j.resmic.2019.07.001. Epub 2019 Jul 17.

DOI:10.1016/j.resmic.2019.07.001
PMID:31325485
Abstract

Bacteria usually produce, release and detect quorum sensing (QS)-based signal molecules, and successively orchestrate gene expression in respond to environmental changes. Pseudoalteromonas are typical marine bacteria, but knowledge on their QS systems is extremely fragmentary. In this study, genome sequencing of Pseudoalteromonas sp. R3 was performed. Accordingly, a QS working model including three sets of hierarchically organized QS systems was proposed in strain R3. Among them, the typical LuxI/R-type QS system using acyl-homoserine lactones (AHLs) as signal molecules was characterized. Sequence similarity analysis indicated luxI encoding AHLs synthase is novel. The luxR encoding AHLs receptor is directly adjacent to luxI downstream. Notably, mutagenesis demonstrated LuxI and LuxR affect each other at transcriptional level, and both control the AHLs formation. Interestingly, it was found that LuxI/R-type QS system positively involves resistance to streptomycin. Thin-layer chromatography analysis showed strain R3 can produce 3-OH-C6-HSL and C8-HSL, which was supported by heterologous expression of LuxI in Escherichia coli. Sequence alignment analysis indicated that the N-terminal region of LuxI is more conservative than the C-terminal region, revealing the importance of N-terminal region in AHLs synthesis. The obtained findings enrich our knowledge on LuxI/R-type QS system in Pseudoalteromonas and its regulation on adaptation to marine environments.

摘要

细菌通常会产生、释放和检测群体感应 (QS) 为基础的信号分子,并依次协调基因表达以响应环境变化。拟交替单胞菌是典型的海洋细菌,但对其 QS 系统的了解非常零碎。在本研究中,对假交替单胞菌 R3 进行了基因组测序。因此,在 R3 菌株中提出了包括三组层次组织的 QS 工作模型。其中,使用酰基高丝氨酸内酯 (AHLs) 作为信号分子的典型 LuxI/R 型 QS 系统得到了表征。序列相似性分析表明,编码 AHLs 合酶的 luxI 是新的。编码 AHLs 受体的 luxR 直接位于 luxI 的下游。值得注意的是,突变分析表明 LuxI 和 LuxR 在转录水平上相互影响,并且两者都控制 AHLs 的形成。有趣的是,发现 LuxI/R 型 QS 系统对链霉素的抗性有积极影响。薄层层析分析表明,R3 菌株可以产生 3-OH-C6-HSL 和 C8-HSL,这得到了在大肠杆菌中异源表达 LuxI 的支持。序列比对分析表明,LuxI 的 N 端区域比 C 端区域更保守,这表明 N 端区域在 AHLs 合成中的重要性。获得的发现丰富了我们对假交替单胞菌中 LuxI/R 型 QS 系统及其对海洋环境适应的调控的认识。

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