目的:抗生素耐药性威胁的上升对公共卫生构成了重大挑战。抗性机制新方向的研究对于克服这一障碍至关重要。本研究通过比较敏感和实验诱导的氧氟沙星耐药大肠杆菌(E。大肠杆菌)使用多组学分析的菌株,旨在为细菌耐药性提供新的见解。
方法:通过暴露于高浓度的氧氟沙星来选择抗氧氟沙星的大肠杆菌菌株。涉及转录组学的比较分析,蛋白质组学,在野生型(WT)和氧氟沙星耐药(Re-OFL)菌株之间进行乙酰组学。差异表达基因的富集途径,使用基因本体论(GO)和京都基因和基因组百科全书(KEGG)方法分析了两种菌株之间的蛋白质和乙酰化蛋白质。此外,使用整合的多组学分析策略对大肠杆菌的代谢网络进行了映射.
结果:我们确定了2775个mRNA的显着差异,1062种蛋白质,和WT和Re-OFL菌株之间的1015个乙酰化蛋白。综合组学分析显示,常见的改变丰富了代谢过程,特别是糖酵解途径。进一步的分析表明,14种代谢酶表现出上调的乙酰化水平和下调的转录和蛋白质水平。此外,这些代谢酶中的七个(FBA,tpi,gapA,pykA,sdhA,FumA,和mdh)是与糖酵解途径相关的成分。
结论:抗生素引起的代谢酶变化似乎是大肠杆菌适应抗生素压力的关键因素,这为理解氧氟沙星压力时的适应机制提供了新的思路。
OBJECTIVE: The rising threat of antibiotic resistance poses a significant challenge to public health. The research on the new direction of resistance mechanisms is crucial for overcoming this hurdle. This study examines metabolic changes by comparing sensitive and experimentally induced ofloxacin-resistant Escherichia coli (E. coli) strains using multi-omics analyses, aiming to provide novel insights into bacterial resistance.
METHODS: An ofloxacin-resistant E. coli strain was selected by being exposed to high concentration of ofloxacin. Comparative analyses involving transcriptomics, proteomics, and acetylomics were conducted between the wild-type (WT) and the ofloxacin-resistant (Re-OFL) strains. Enrichment pathways of differentially expressed genes, proteins and acetylated proteins between the two strains were analyzed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) method. In addition, the metabolic network of E. coli was mapped using integrated multi-omics analysis strategies.
RESULTS: We identified significant differences in 2775 mRNAs, 1062 proteins, and 1015 acetylated proteins between WT and Re-OFL strains. Integrated omics analyses revealed that the common alterations enriched in metabolic processes, particularly the glycolytic pathway. Further analyses demonstrated that 14 metabolic enzymes exhibited upregulated acetylation levels and downregulated transcription and protein levels. Moreover, seven of these metabolic enzymes (fba, tpi, gapA, pykA, sdhA, fumA, and mdh) were components related to the glycolytic pathway.
CONCLUSIONS: The changes of metabolic enzymes induced by antibiotics seem to be a key factor for E. coli to adapt to the pressure of antibiotics, which shed new light on understanding the adaptation mechanism when responding to ofloxacin pressure.