The activity of the quorum sensing regulator HapR is modulated by the bacterial extracellular vesicle (BEV)-associated protein ObfA of Vibrio cholerae

IF 15.5 1区 医学 Q1 CELL BIOLOGY
Stephan P. Ebenberger, Fatih Cakar, Yi-Chi Chen, Katharina Pressler, Leo Eberl, Stefan Schild
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引用次数: 0

Abstract

Vibrio cholerae, a facultative human pathogen and causative agent of the severe diarrheal disease cholera, transits between the human intestinal tract and aquatic reservoirs. Like other bacterial species, V. cholerae continuously releases bacterial extracellular vesicles (BEVs) from its surface, which have been recently characterised for their role during in vivo colonisation. However, between epidemic outbreaks, V. cholerae persists in the biofilm mode for extended periods in aquatic reservoirs, which enhances environmental fitness and host transition. In this study, we investigated the effect of V. cholerae BEVs on biofilm formation, a critical feature for ex vivo survival. In contrast to BEVs from planktonic cultures, our results show that physiological concentrations of BEVs from dynamic biofilm cultures facilitate V. cholerae biofilm formation, which could be linked to a proteinaceous factor. Comparative proteomic analyses of planktonic- and biofilm-derived BEVs identified a previously uncharacterised outer membrane protein as an abundant component of dynamic biofilm-derived BEVs, which was found to be responsible for the BEV-dependent enhancement of biofilm production. Consequently, this protein was named outer membrane-associated biofilm facilitating protein A (ObfA). Comprehensive molecular studies unravelled ObfA as a negative modulator of HapR activity. HapR is a key transcriptional regulator of the V. cholerae quorum sensing (QS) cascade acting as a potent repressor of biofilm formation and virulence. Consistently, obfA mutants not only exhibited reduced biofilm production but also reduced colonisation fitness. Surprisingly, our results demonstrate that ObfA does not affect HapR through the canonical QS system but via the Csr-cascade altering the expression of the small regulatory RNAs CsrC and CsrD. In summary, this study elucidates a novel intraspecies BEV-based communication in V. cholerae that influences biofilm formation and colonisation fitness via a new regulatory pathway involving HapR, Csr-cascade and the BEV-associated protein ObfA.

Abstract Image

霍乱弧菌的细菌胞外囊泡(BEV)相关蛋白 ObfA 可调节法定人数感应调节因子 HapR 的活性
霍乱弧菌是一种面性人类病原体,也是严重腹泻病霍乱的致病菌,它在人类肠道和水生水库之间传播。与其他细菌物种一样,霍乱弧菌不断从其表面释放细菌胞外囊泡(BEVs),这些囊泡在体内定植过程中的作用最近已得到证实。然而,在流行病爆发的间歇期,霍乱弧菌会在水生水库中长时间以生物膜模式存活,从而提高环境适应性和宿主转换能力。在这项研究中,我们调查了霍乱弧菌 BEV 对生物膜形成的影响,生物膜是霍乱弧菌体内外存活的关键特征。与来自浮游生物培养物的 BEVs 不同,我们的研究结果表明,来自动态生物膜培养物的生理浓度的 BEVs 会促进霍乱弧菌生物膜的形成,这可能与蛋白质因子有关。对浮游生物和生物膜衍生的 BEV 进行的蛋白质组学比较分析发现,动态生物膜衍生的 BEV 中含有一种以前未定性的外膜蛋白,它是 BEV 依赖性增强生物膜生成的原因。因此,这种蛋白质被命名为外膜相关生物膜促进蛋白 A(ObfA)。综合分子研究发现,ObfA 是 HapR 活性的负调制剂。HapR 是霍乱弧菌法定量感应(QS)级联的一个关键转录调节因子,对生物膜的形成和毒力起着强有力的抑制作用。一致的是,obfA 突变体不仅生物膜生成减少,而且定殖能力也降低。令人惊讶的是,我们的研究结果表明,ObfA 并不是通过典型的 QS 系统影响 HapR,而是通过 Csr 级联改变小调控 RNA CsrC 和 CsrD 的表达。总之,本研究阐明了霍乱弧菌中一种新的种内基于 BEV 的交流,这种交流通过涉及 HapR、Csr 级联和 BEV 相关蛋白 ObfA 的新调控途径影响生物膜的形成和定殖适应性。
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来源期刊
Journal of Extracellular Vesicles
Journal of Extracellular Vesicles Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
27.30
自引率
4.40%
发文量
115
审稿时长
12 weeks
期刊介绍: The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies. The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.
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