Research Article: LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

Date Published: October 10, 2017

Publisher: Springer Berlin Heidelberg

Author(s): Yuxi Zhang, Kezong Qi, Yawei Jing, Jiakun Zuo, Jiangang Hu, Xiaolong Lv, Zhaoguo Chen, Rongsheng Mi, Yan Huang, Shengqing Yu, Xiangan Han.

http://doi.org/10.1186/s13568-017-0486-y

Abstract

The luxS gene is required for autoinducer-2 (AI-2) synthesis in many bacterial species. AI-2 is taken up by a specific receptor to regulate multiple bacterial activities. However, the lack of methods to identify AI-2 receptors has impeded investigations into the roles of AI-2. Here, a luxS mutant of Escherichia coli strain BL21 (DE3) was constructed (named BL21∆luxS), and the recombinant LsrB protein of Salmonella enterica was expressed in BL21∆luxS and BL21 cells, which were named LsrB (BL21∆luxS) and LsrB (BL21), respectively. The results of the activity of recombinant LsrB binding showed that LsrB (BL21) bound to endogenous AI-2 (produced from BL21 strain), while LsrB (BL21∆luxS) did not (as BL21∆luxS cannot produce AI-2). However, the results of recombinant LsrB binding showed that LsrB (BL21∆luxS) can bind exogenous AI-2, which was released from LsrB (BL21∆luxS) at 55 °C for 10 min, while LsrB (BL21) could not bind exogenous AI-2 (due to binding of endogenous AI-2 before). Furthermore, analysis of the thermal stability of AI-2 showed that that AI-2 activity was relatively high at incubation temperatures below 65 °C. These findings will be beneficial for screening of new AI-2 receptors in different bacterial species.

Partial Text

Cell–cell communication in bacteria is accomplished through the exchange of extracellular signalling molecules, called autoinducers, in a process termed quorum sensing (QS) (Pereira et al. 2013). Although most autoinducers are species specific, autoinducer-2 (AI-2) is considered to be a universal signalling molecule for interspecies communication that is synthesised by LuxS, an enzyme that is highly conserved and widespread in diverse bacteria (Han and Lu 2009; Pereira et al. 2013; Even-Tov et al. 2016).

AI-2, which was first identified in the marine bacterium V. harveyi, is produced by many Gram-negative and -positive bacteria (Surette et al. 1999). AI-2 is produced by the activated methyl cycle by the AI-2 synthase Pfs and LuxS. The LuxS-mediated QS system play roles in biofilm formation, virulence gene expression, the type III secretion system, and drug sensitivity changes in different bacterial species by AI-2 (Xue et al. 2013; Novotny et al. 2015; Mou and Plummer 2016). AI-2 is taken up by specific membrane receptors in bacterial transport systems to regulate physiological functions, such as LuxP in V. harveyi (Chen et al. 2002), LsrB in S. typhimurium (Miller et al. 2004) and RbsB in H. influenza (Armbruster et al. 2011). Furthermore, the AI-2 receptors also play roles in some host-microbial relationships. For example, an AI-2 mimic detected by the bacterial AI-2 receptor, LuxP/LsrB, can activate QS-controlled gene expression in the enteric pathogen S. typhimurium (Ismail et al. 2016). Therefore, further research of the AI-2 receptors will help to elucidate the mechanisms underlying the LuxS/AI-2 type QS system.

 

Source:

http://doi.org/10.1186/s13568-017-0486-y

 

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