The Global Acetylome of the Human Pathogen Vibrio cholerae V52 Reveals Lysine Acetylation of Major Transcriptional Regulators

Abstract

Protein lysine acetylation is recognized as an important reversible post translational modification in all domains of life. While its primary roles appear to reside in metabolic processes, lysine acetylation has also been implicated in regulating pathogenesis in bacteria. Several global lysine acetylome analyses have been carried out in various bacteria, but thus far there have been no reports of lysine acetylation taking place in the important human pathogen Vibrio cholerae. In this study, we analyzed the lysine acetylproteome of the human pathogen V. cholerae V52. By applying a combination of immuno-enrichment of acetylated peptides and high resolution mass spectrometry, we identified 3,402 acetylation sites on 1,240 proteins. Of the acetylated proteins, more than half were acetylated on two or more sites. As reported for other bacteria, we observed that many of the acetylated proteins were involved in metabolic and cellular processes and there was an over-representation of acetylated proteins involved in protein synthesis. Of interest, we demonstrated that many global transcription factors such as CRP, H-NS, IHF, Lrp and RpoN as well as transcription factors AphB, TcpP, and PhoB involved in direct regulation of virulence in V. cholerae were acetylated. In conclusion, this is the first global protein lysine acetylome analysis of V. cholerae and should constitute a valuable resource for in-depth studies of the impact of lysine acetylation in pathogenesis and other cellular processes.

A Putative Acetylation System in Vibrio cholerae Modulates Virulence in Arthropod Hosts

“…However, V. cholerae SIO lacks the ctxAB and tcp gene clusters, and virulence in this strain appears to be entirely contingent upon the regulation of metabolite consumption. Intriguingly, Acs in V. cholerae strain V52 is acetylated, but not at the conserved lysine, as in Salmonella and E. coli (39). This observation is consistent with our own results in which mutation of Acs K609 or Acs L641 , when introduced into a ΔcobB background, did not improve acetate metabolism or virulence in vivo, suggesting that these mutations could not prevent acetylation.…”

“…Acetylation modifies a sizeable proportion of the proteome in a number of bacterial pathogens, including Vibrio parahaemolyticus, Mycobacterium tuberculosis, Staphylococcus aureus, and V. cholerae, but these modifications have been directly linked to virulence only in Salmonella and E. coli (38,39). Acetylation regulates the expression of Salmonella type III secretion system 1 by controlling the stability and DNA binding capability of a transcriptional regulator (40,41).…”

“…A very recent study reported that the proteome of V. cholerae V52, a strain pathogenic to humans, is subject to acetylation (39). A number of global regulators important for both virulence and metabolic regulation are acetylated.…”

Lysine acetylation contributes to development, aflatoxin biosynthesis and pathogenicity in Aspergillus flavus

“…The prior system‐wide studies involving lysine‐acetylated peptide immunoprecipitation and liquid chromatography–mass spectrometry/mass spectrometry (LC–MS/MS) measurements was first used in bacterial acetylomes of Escherichia coli (Byung Jo et al ., ). The same approach has since been employed to analyse the other prokaryotic model organisms, including Bacillus subtilis (Kim et al ., ), Mycobacterium tuberculosis (Liu et al ., ), Vibrio cholerae (Jers et al ., ) and Streptococcus pneumoniae (Liu et al ., ). In addition, the acetylome of fungi such as Saccharomyces cerevisiae (Henriksen et al ., ), Candida albicans (Zhou et al ., ), Beauveria bassiana (Wang et al ., ), Botrytis cinerea (Lv et al ., ), Fusarium graminearum (Zhou et al ., ) and Histoplasma capsulatum (Xie et al ., ) have also been reported, and most acetylated proteins are found to play crucial roles in central metabolism, protein synthesis, cell cycle and virulence.…”

Removal of a Membrane Anchor Reveals the Opposing Regulatory Functions of Vibrio cholerae Glucose-Specific Enzyme IIA in Biofilms and the Mammalian Intestine

“…In addition, a recent comprehensive analysis of acetylation sites in the V. cholerae proteome suggests that the lysine at position 6 within the AH is acetylated (38). We predict that acetylation of the EIIA Glc AH should also modulate membrane association.…”