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严格反应因子 DksA 有助于脂肪酸降解功能,从而影响细胞膜稳定性和多黏菌素 B 耐药性。

Stringent Response Factor DksA Contributes to Fatty Acid Degradation Function to Influence Cell Membrane Stability and Polymyxin B Resistance of .

机构信息

Beijing Laboratory for Food Quality and Safety, College of Food Science & Nutritional Engineering, China Agricultural University, 17 Qinghua East Rd., Beijing 100083, China.

出版信息

Int J Mol Sci. 2023 Jul 26;24(15):11951. doi: 10.3390/ijms241511951.

Abstract

DksA is a proteobacterial regulator that binds directly to the secondary channel of RNA polymerase with (p)ppGpp and is responsible for various bacterial physiological activities. While (p)ppGpp is known to be involved in the regulation and response of fatty acid metabolism pathways in many foodborne pathogens, the role of DksA in this process has yet to be clarified. This study aimed to characterize the function of DksA on fatty acid metabolism and cell membrane structure in . Therefore, comparison analysis of gene expression, growth conditions, and membrane permeabilization among the wide-type (WT), DksA-deficient mutant (YEND), and the complemented strain was carried out. It confirmed that deletion of DksA led to a more than four-fold decrease in the expression of fatty acid degradation genes, including . Additionally, YEND exhibited a smaller growth gap compared to the WT strain at low temperatures, indicating that DksA is not required for the growth of in cold environments. Given that polymyxin B is a cationic antimicrobial peptide that targets the cell membrane, the roles of DksA under polymyxin B exposure were also characterized. It was found that DksA positively regulates the integrity of the inner and outer membranes of under polymyxin B, preventing the leakage of intracellular nucleic acids and proteins and ultimately reducing the sensitivity of to polymyxin B. Taken together, this study provides insights into the functions of DksA and paves the way for novel fungicide development.

摘要

DksA 是一种细菌调节因子,可直接与 RNA 聚合酶的次要通道结合,并与 (p)ppGpp 结合,负责各种细菌生理活动。虽然 (p)ppGpp 已知参与许多食源性病原体中脂肪酸代谢途径的调节和反应,但 DksA 在该过程中的作用尚未阐明。本研究旨在研究 DksA 在脂肪酸代谢和细胞膜结构中的功能。因此,对野生型 (WT)、DksA 缺陷突变体 (YEND) 和互补菌株的基因表达、生长条件和膜通透性进行了比较分析。结果证实,DksA 的缺失导致脂肪酸降解基因的表达下降了四倍以上,包括。此外,与 WT 菌株相比,YEND 在低温下的生长差距较小,表明 DksA 不是低温下生长的必需条件。鉴于多粘菌素 B 是一种靶向细胞膜的阳离子抗菌肽,还对多粘菌素 B 暴露下 DksA 的作用进行了表征。结果发现,DksA 可正向调节多粘菌素 B 下 的内外膜的完整性,防止细胞内核酸和蛋白质的泄漏,最终降低 对多粘菌素 B 的敏感性。总之,本研究深入了解了 DksA 的功能,并为新型杀菌剂的开发铺平了道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cbd/10418728/1c9f984ec3a1/ijms-24-11951-g001.jpg

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