Yang Yang, Wu Ziyan, Zhu Li'e, Han Zixin, Li Junpeng, Fang Qiaoqiao, Zhu Guoqiang
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Jiangsu Co-Innovation Center for Important Animal Infectious Diseases and Zoonoses, Joint Laboratory of International Cooperation on Prevention and Control Technology of Important Animal Diseases and Zoonoses of Jiangsu, Yangzhou 225009, China.
Microorganisms. 2025 Aug 6;13(8):1837. doi: 10.3390/microorganisms13081837.
Antibiotic misuse accelerates resistance dissemination via plasmid conjugation, but quorum sensing (QS) regulatory mechanisms remain undefined. Using () MG1655 conjugation models (RP4-7/EC600 plasmids), we demonstrate that long-chain acyl-homoserine lactones (C10/C12-HSL) enhance transfer frequency by up to 7.7-fold (200 μM C12-HSL; < 0.001), while quorum-quenching by sub-inhibitory vanillin suppressed this effect by 95% ( < 0.0001). C12-HSL compromised membrane integrity via upregulation (4-fold; < 0.01) and conjugative pore assembly ( upregulated by 1.38-fold; < 0.05), coinciding with ROS accumulation (1.5-fold; < 0.0001) and SOS response activation ( upregulated by 1.68-fold; < 0.001). Crucially, and deletion mutants reduced conjugation by 65.5% and 55.8%, respectively ( < 0.001), exhibiting attenuated membrane permeability (≤65.5% reduced NPN influx; < 0.0001), suppressed ROS (≤54% downregulated; < 0.0001), and abolished transcriptional induction of conjugation/stress genes. Reciprocal RpoS-RMF (ribosomal hibernation factor) crosstalk was essential for AHL responsiveness, with deletions mutually suppressing expression (≤65.9% downregulated; < 0.05). We establish a hierarchical mechanism wherein long-chain AHLs drive resistance dissemination through integrated membrane restructuring, stress adaptation, and RpoS-RMF-mediated genetic plasticity, positioning QS signaling as a viable target for curbing resistance spread.
抗生素的滥用通过质粒接合加速了耐药性的传播，但群体感应（QS）调控机制仍不明确。利用（）MG1655接合模型（RP4 - 7/EC600质粒），我们证明长链酰基高丝氨酸内酯（C10/C12 - HSL）可将转移频率提高多达7.7倍（200 μM C12 - HSL；<0.001），而亚抑制浓度香草醛的群体猝灭作用可将这种效应抑制95%（<0.0001）。C12 - HSL通过上调（4倍；<0.01）和接合孔组装（上调1.38倍；<0.05）破坏膜完整性，这与活性氧积累（1.5倍；<0.0001）和SOS反应激活（上调1.68倍；<0.001）同时发生。至关重要的是， 和 缺失突变体分别使接合减少了65.5%和55.8%（<0.001），表现出膜通透性减弱（NPN内流减少≤65.5%；<0.0001）、活性氧受抑制（下调≤54%；<0.0001）以及接合/应激基因的转录诱导被消除。RpoS与核糖体休眠因子（RMF）的相互作用对于AHL反应性至关重要，缺失会相互抑制表达（下调≤65.9%；<0.05）。我们建立了一种分级机制，其中长链AHL通过整合膜重塑、应激适应和RpoS - RMF介导的遗传可塑性来驱动耐药性传播，将QS信号定位为遏制耐药性传播的可行靶点。
