Genome- and Proteome-Wide Analysis of Lysine Acetylation in Vibrio vulnificus Vv180806 Reveals Its Regulatory Roles in Virulence and Antibiotic Resistance

Abstract

Infection with Vibrio vulnificus is notorious for its atypical clinical manifestations and irreversible disease progression. Lysine acetylation is a conserved post-translational modification (PTM) that plays a critical regulatory role in diverse cellular processes. However, little is known about the role of lysine acetylation on the pathogenesis of V. vulnificus. Here, we report the complete genome sequence and a global profile for protein lysine acetylation of V. vulnificus Vv180806, a highly cefoxitin resistant strain isolated from a mortality case. The assembled genome comprised two circular chromosomes and one circular plasmid; it contained 4,770 protein-coding genes and 153 RNA genes. Phylogenetic analysis revealed genetic homology of this strain with other V. vulnificus strains from food sources. Of all the proteins in this strain, 1,924 (40.34%) were identified to be acetylated at 6,626 sites. The acetylated proteins were enriched in metabolic processes, binding functions, cytoplasm, and multiple central metabolic pathways. Moreover, the acetylation was found in most identified virulence factors of this strain, suggesting its potentially important role in bacterial virulence. Our work provides insights into the genomic and acetylomic features responsible for the virulence and antibiotic resistance of V. vulnificus, which will facilitate future investigations on the pathogenesis of this bacterium.

A Comprehensive Analysis of the Lysine Acetylome in the Aquatic Animals Pathogenic Bacterium Vibrio mimicus

“…This has also been discovered in diverse organisms, which may make clear that acetylation plays a jointly regulatory role in central metabolic enzymes ( Choudhary et al, 2014 ). Furthermore, previous research reported that large-scale ribosome-related proteins were acetylated, such as in A. hydrophila , V. vulnificus , and S. pneumoniae ; our results also indicated the proteins involved in ribosome were more abundant through KEGG enrichment analysis ( Liu et al, 2018 ; Sun et al, 2019 ; Pang et al, 2020 ).…”

“…MF analysis revealed that acetylated proteins were outstandingly enriched in binding, heterocyclic compound binding, organic cyclic compound binding, and ion binding ( Figure 3A ). Consistent with previous research, the identified acetylproteins mainly enrich for metabolism processes, location of the cell, and binding function, which are especially similar to Vibrio spp., such as Vibrio cholerae V52, Vibrio alginolyticus , Vibrio parahemolyticus , and Vibrio vulnificus Vv180806 ( Pan et al, 2014 ; Carsten et al, 2017 ; Pang et al, 2019 ; Pang et al, 2020 ).…”

“…The target proteins were precipitated with specific polyclonal antibodies against LuxO and LuxR. V. mimicus strain whole protein (500 μg) was incubated with LuxO and LuxR antibody at 4°C overnight, as previously described ( Pang et al, 2020 ). Briefly, the protein A/G beads were washed three times with PBS buffer and then added to the lysate for incubation at 4°C for 1–3 h. The beads were centrifuged at 4°C and 3,000 g for 5 min to remove the supernatant, and washed five times with PBS buffer.…”

“…The transfer of acetyl group to the side chain of lysine is realized by enzymatic or non-enzymatic, and the enzymatic reaction is realized with acetyl phosphate or acetyl coenzyme a as the cofactor of lysine acetyltransferase (KAT) ( Carabetta and Cristea, 2017 ; Macek et al, 2019 ). At present, more and more studies on lysine acylated proteins have been reported in various bacterial pathogens, such as Porphyromonas gingivalis ( Butler et al, 2015 ), Pseudomonas aeruginosa ( Ouidir et al, 2015 ), Aeromonas hydrophila ( Sun et al, 2019 ), Vibrio cholerae ( Carsten et al, 2017 ), Vibrio vulnificus ( Pang et al, 2020 ), and so on. In addition, acetylation modification occurs on enzymes in several central metabolic processes of bacteria, such as tricarboxylic acid cycle, glycolysis/gluconeogenesis, and pyruvate metabolism ( Li W.X.…”

Contribution of N ε -lysine Acetylation towards Regulation of Bacterial Pathogenesis

“…However, none of these predictions have been experimentally confirmed; therefore, the physiologic significance of these observations remains unclear. The acetylomes of other pathogenic Vibrio species have also been characterized, including V. vulnificus ( 37 ), V. parahaemolyticus ( 38 ), and V. alginolyticus ( 39 ), but investigation of potential virulence factors was not extensively explored.…”