Dataset Information

Global acetylomics of Campylobacter jejuni shows lysine acetylation regulates CadF adhesin processing and fibronectin binding during in vivo-like growth

ABSTRACT: Campylobacter jejuni is the leading cause of acute bacterial gastroenteritis in the developed world. Despite the prevalence of infection, the pathogenesis of C. jejuni remains poorly understood. Lysine acetylation (KAc) is a reversible post-translational modification (PTM) that can alter the protein structure / function paradigm, however specific roles for KAc are largely undefined in bacteria. Acetyl-lysine immunoprecipitation and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) identified 5567 acetylated lysines on 1026 C. jejuni proteins (~63% of the predicted C. jejuni proteome). Functional enrichment confirmed acetylated proteins are involved in metabolism, transcription and translation, stress responses and chemotaxis. KAc was identified on proteins from all subcellular locations, including outer membrane (OM) and extracellular proteins. Label-based LC-MS/MS identified acetylated proteins and modified sites associated with growth in 0.1% sodium deoxycholate (DOC, a component of gut bile salts). A total of 3571 acetylated peptides were quantified and 761 (from 409 proteins) were differentially abundant following growth in DOC. Changes in KAc involved multiple pathways suggesting a dynamic role for this PTM in bile resistance. As observed in other bacteria, we show that KAc is primarily non-enzymatically mediated via acetyl-phosphate; however, the sole deacetylase CobB also contributes to a global elevation of this modification in DOC. We observed several multiply acetylated OM proteins and altered DOC abundance of acetylated peptides in the major fibronectin (Fn)-binding adhesin CadF. We show that KAc influences CadF Fn binding and prevalence of processed lower mass variants. This study provides the first system-wide analysis of the C. jejuni lysine acetylome and contributes to our understanding of KAc as an emerging PTM in bacteria.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Campylobacter Jejuni Subsp. Jejuni Nctc 11168

TISSUE(S): Cell Culture

SUBMITTER: Ashleigh Dale

LAB HEAD: Stuart James Cordwell

PROVIDER: PXD042815 | Pride | 2025-03-11

REPOSITORIES: Pride

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			Action	DRS
	20200324_QEHFX3_MYW_AD_KAcUnbound_Fr1.raw	Raw
	20200324_QEHFX3_MYW_AD_KAcUnbound_Fr10.raw	Raw
	20200324_QEHFX3_MYW_AD_KAcUnbound_Fr11.raw	Raw
	20200324_QEHFX3_MYW_AD_KAcUnbound_Fr12.raw	Raw
	20200324_QEHFX3_MYW_AD_KAcUnbound_Fr13.raw	Raw

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Global Acetylomics of <i>Campylobacter jejuni</i> Shows Lysine Acetylation Regulates CadF Adhesin Processing and Human Fibronectin Binding.

Dale Ashleigh L AL Man Lok L Cordwell Stuart J SJ

Journal of proteome research 20231013 11

Lysine acetylation (KAc) is a reversible post-translational modification (PTM) that can alter protein structure and function; however, specific roles for KAc are largely undefined in bacteria. Acetyl-lysine immunoprecipitation and LC-MS/MS identified 5567 acetylated lysines on 1026 proteins from the gastrointestinal pathogen <i>Campylobacter jejuni</i> (∼63% of the predicted proteome). KAc was identified on proteins from all subcellular locations, including the outer membrane (OM) and extracellu ...[more]

PMID: 37830485

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Project description:C. jejuni, a spiral-shaped gram-negative bacterium, is a leading bacterial cause of human foodborne illness. Acute disease is associated with C. jejuni invasion of the intestinal epithelium. Further, maximal host cell invasion requires the secretion of proteins termed Campylobacter invasion antigens (Cia). As bile acids are known to alter the pathogenic behavior of other gastrointestinal pathogens, we hypothesized that the virulence potential of Campylobacter may be triggered by the bile acid deoxycholate (DOC). In support of this hypothesis, culturing C. jejuni with a physiologically relevant concentration of DOC significantly altered the kinetics of cell invasion as evidenced by gentamicin-protection assays. In contrast to C. jejuni harvested from Mueller-Hinton (MH) agar plates, C. jejuni harvested from MH agar plates supplemented with DOC demonstrated Cia secretion as judged by metabolic labeling experiments. DOC was also found to induce the expression of the ciaB gene as judged by B-galactosidase reporter assays and real-time RT-PCR. Microarray analysis revealed that DOC induced the expression of virulence genes (i.e., ciaB, cmeABC, dccR, and tlyA). In summary, we demonstrate that it is possible to enhance the pathogenic behavior of C. jejuni by modifying the culture conditions. These results provide a foundation to identify genes expressed by C. jejuni in response to in vivo-like culture conditions. Keywords: Stress response For the expression profiling arrays, an indirect comparison of gene expression was performed, where the expression profile of the C. jejuni F38011 cultured in the presence and absence of DOC was measured separately on different slides as described previously (26). Briefly, Cy5 labeled reference DNA from the C. jejuni F38011 strain was mixed with Cy3 labeled test cDNA (C. jejuni F38011 cultured in the presence or absence of DOC) and hybridized to the Campylobacter cDNA array (26) on separate slides. DNA microarrays were scanned using an Axon GenePix 4000B microarray laser scanner (Axon Instruments, Union City, CA) and the data for spot and background intensities were processed using the GenePix 4.0 software. To compensate for any effect of the amount of template and uneven Cy-dye incorporation, data normalization was performed as previously described (26). For the comparison of genes differentially expressed in the presence and absence of DOC,six hybridization measurements were generated per biological experiment (three technical replicate arrays and two replicate features per array).