Lysine Acetylation Regulates Antibiotic Resistance In Escherichia Coli

Antibiotic resistance is increasingly becoming a serious challenge to public health.The regulation of metabolism by post-translational modifications(PTMs)has been widely studied;however,the comprehensive mechanism underlying the regulation of acetylation in bacterial resistance against antibiotics is unknown.Herein,with Escherichia coli as the study model,we performed quantitative analysis of the acetylated proteome of wild-type strain(WT)and ampicillin-(Re-Amp),kanamycin-(Re-Kan),and polymyxin B-resistant(Re-Pol)strains.Based on bioinformatics analysis combined with biochemical validations,we aimed to find a possible common regulatory mechanism existing between the different resistant strains.The results showed that acetylation negatively regulates bacterial metabolism to maintain antibiotic resistance,but positively regulates bacterial motility.Further analyses revealed that key enzymes in various metabolic pathways were differentially acetylated.Particularly,pyruvate kinase(PykF),a key glycolytic enzyme regulating bacterial metabolism,and its acetylated form were highly expressed in the three resistant types and were identified as reversibly acetylated by the deacetylase Cob B and the acetyl-transferase Pat Z,and also could be acetylated by non-enzyme Ac P in vitro.Further,the deacetylation of Lys413 of PykF increased the enzyme activity by changing the conformation of ATP binding site of PykF,resulting in an increase in energy production,which in turn increased the sensitivity of drug-resistant strains to antibiotics.This study provides novel insights for understanding bacterial resistance and lays the foundation for future research on regulation of acetylation in antibiotic-resistant strains.