First acetyl-proteome profiling of Salmonella Typhimurium revealed involvement of lysine acetylation in drug resistance

Abstract

Salmonella are becoming increasingly resistant to fluoroquinolones (FQs), therefore determining the resistance mechanism is very important. Recent studies have shown that protein post-translational modifications (PTM) play a role in bacterial antibiotic resistance. One such type of PTM, lysine acetylation, is a reversible and highly regulated PTM which has been found to be associated with antibiotic resistance in Mycobacterium and Acinetobacter species. Salmonella Typhimurium are major zoonotic pathogens, which are becoming increasingly resistant to FQs, the antibiotics of choice where therapy is indicated. To date, however, there have been no studies on the relationship between PTM and drug resistance in Salmonella. Therefore, in the present study, ciprofloxacin-resistant and susceptible strains of Salmonella were used as the research objects, and tandem mass tag labeling and acetylation enrichment techniques were used to screen for the different expression of actylated proteins between the two strains, and for quantitative and bioinformatics analysis. We identified a total of 631 acetylated proteins involving 1259 lysine acetylation sites. Among the quantified sites, compared with the susceptible strain, the expression of lysine acetylation was upregulated for 112 sites and downregulated for 149 sites in the resistant strain. Bioinformatic analyses showed that the main enrichment pathways for these differentially acetylated proteins are microbial metabolic process, biosynthesis of antibiotics, and bacterial chemotaxis. Among the differentially acetylated proteins, 14 proteins related to bacterial antibiotic resistance were identified (excluding metabolic and virulence-related proteins), and the lysine acetylation expression of these proteins was significantly different between the resistant and susceptible strains. These results indicated that protein lysine acetylation is not only related to metabolism and virulence, but also to antibiotic resistance. The results provide an important basis for in-depth studies of the relationship between protein lysine acetylation and bacterial antibiotic resistance.

Keywords: Acetylation; Bacterial antibiotic resistance; Protein post-translational modification; Salmonella.