Campylobacter jejuni carbon starvation protein A (CstA) is involved in peptide utilization, motility and agglutination, and has a role in stimulation of dendritic cells
Campylobacter jejuni is the most frequent cause of severe gastroenteritis in the developed world. The major symptom of campylobacteriosis is inflammatory diarrhoea. The molecular mechanisms of this infection are poorly understood compared to those of less frequent disease-causing pathogens. In a previous study, we identified C. jejuni proteins that antibodies in human campylobacteriosis patients reacted with. One of the immunogenic proteins identified (Cj0917) displays homology to carbon starvation protein A (CstA) from Escherichia coli, where this protein is involved in the starvation response and peptide uptake. In contrast to many bacteria, C. jejuni relies on amino acids and organic acids for energy, but in vivo it is highly likely that peptides are also utilized, although their mechanisms of uptake are unknown. In this study, Biolog phenotype microarrays have been used to show that a ΔcstA mutant has a reduced ability to utilize a number of di- and tri-peptides as nitrogen sources. This phenotype was restored through genetic complementation, suggesting CstA is a peptide uptake system in C. jejuni. Furthermore, the ΔcstA mutant also displayed reduced motility and reduced agglutination compared to WT bacteria; these phenotypes were also restored through complementation. Murine dendritic cells exposed to UV-killed bacteria showed a reduced IL-12 production, but the same IL-10 response when encountering C. jejuni ΔcstA compared to the WT strain. The greater Th1 stimulation elicited by the WT as compared to ΔcstA mutant cells indicates an altered antigenic presentation on the surface, and thus an altered recognition of the mutant. Thus, we conclude that C. jejuni CstA is important not only for peptide utilization, but also it may influence host–pathogen interactions.
Andersen-NissenE., SmithK. D., StrobeK. L., BarrettS. L., CooksonB. T., LoganS. M., AderemA.2005; Evasion of Toll-like receptor 5 by flagellated bacteria. Proc Natl Acad Sci U S A 102:9247–9252 [View Article][PubMed]
BaxM., KuijfM. L., HeikemaA. P., van RijsW., BruijnsS. C. M., García-VallejoJ. J., CrockerP. R., JacobsB. C., van VlietS. J., van KooykY.2011; Campylobacter jejuni lipooligosaccharides modulate dendritic cell-mediated T cell polarization in a sialic acid linkage-dependent manner. Infect Immun 79:2681–2689 [View Article][PubMed]
BlumP. H., JovanovichS. B., McCannM. P., SchultzJ. E., LesleyS. A., BurgessR. R., MatinA.1990; Cloning and in vivo and in vitro regulation of cyclic AMP-dependent carbon starvation genes from Escherichia coli.. J Bacteriol 172:3813–3820[PubMed]
CohnM. T., IngmerH., MulhollandF., JørgensenK., WellsJ. M., BrøndstedL.2007; Contribution of conserved ATP-dependent proteases of Campylobacter jejuni to stress tolerance and virulence. Appl Environ Microbiol 73:7803–7813 [View Article][PubMed]
de ZoeteM. R., KeestraA. M., WagenaarJ. A., van PuttenJ. P. M.2010; Reconstitution of a functional Toll-like receptor 5 binding site in Campylobacter jejuni flagellin. J Biol Chem 285:12149–12158 [View Article][PubMed]
FryB. N., FengS., ChenY. Y., NewellD. G., ColoeP. J., KorolikV.2000; The galE gene of Campylobacter jejuni is involved in lipopolysaccharide synthesis and virulence. Infect Immun 68:2594–2601 [View Article][PubMed]
GoldenN. J., AchesonD. W. K.2002; Identification of motility and autoagglutination Campylobacter jejuni mutants by random transposon mutagenesis. Infect Immun 70:1761–1771 [View Article][PubMed]
GoossensH., VlaesL., GalandI., Van den BorreC., ButzlerJ.-P.1989; Semisolid blood-free selective-motility medium for the isolation of campylobacters from stool specimens. J Clin Microbiol 27:1077–1080[PubMed]
GrantC. C. R., KonkelM. E., CieplakW.Jr, TompkinsL. S.1993; Role of flagella in adherence, internalization, and translocation of Campylobacter jejuni in nonpolarized and polarized epithelial cell cultures. Infect Immun 61:1764–1771[PubMed]
GrippE., HlahlaD., DidelotX., KopsF., MaurischatS., TedinK., AlterT., EllerbroekL., SchreiberK.other authors2011; Closely related Campylobacter jejuni strains from different sources reveal a generalist rather than a specialist lifestyle. BMC Genomics 12:584 [View Article][PubMed]
GroatR. G., SchultzJ. E., ZychlinskyE., BockmanA., MatinA.1986; Starvation proteins in Escherichia coli: kinetics of synthesis and role in starvation survival. J Bacteriol 168:486–493[PubMed]
GuccioneE., Leon-KempisM. R., PearsonB. M., HitchinE., MulhollandF., van DiemenP. M., StevensM. P., KellyD. J.2008; Amino acid-dependent growth of Campylobacter jejuni: key roles for aspartase (AspA) under microaerobic and oxygen-limited conditions and identification of AspB (Cj0762), essential for growth on glutamate. Mol Microbiol 69:77–93 [View Article][PubMed]
HoltJ. P., GrantA. J., CowardC., MaskellD. J., QuinlanJ. J.2012; Identification of Cj1051c as a major determinant for the restriction barrier of Campylobacter jejuni strain NCTC11168. Appl Environ Microbiol 78:7841–7848 [View Article][PubMed]
HowlettR. M., HughesB. M., HitchcockA., KellyD. J.2012; Hydrogenase activity in the foodborne pathogen Campylobacter jejuni depends upon a novel ABC-type nickel transporter (NikZYXWV) and is SlyD-independent. Microbiology 158:1645–1655 [View Article][PubMed]
KarlyshevA. V., WrenB. W.2005; Development and application of an insertional system for gene delivery and expression in Campylobacter jejuni.. Appl Environ Microbiol 71:4004–4013 [View Article][PubMed]
KuijfM. L., SamsomJ. N., van RijsW., BaxM., HuizingaR., HeikemaA. P., van DoornP. A., van BelkumA., van KooykY.other authors2010; TLR4-mediated sensing of Campylobacter jejuni by dendritic cells is determined by sialylation. J Immunol 185:748–755 [View Article][PubMed]
Leon-KempisM., GuccioneE. J., MulhollandF., WilliamsonM. P., KellyD. J.2006; The Campylobacter jejuni PEB1a adhesin is an aspartate/glutamate-binding protein of an ABC transporter essential for microaerobic growth on dicarboxylic amino acids. Mol Microbiol 60:1262–1275 [View Article][PubMed]
ParkhillJ., WrenB. W., MungallK., KetleyJ. M., ChurcherC., BashamD., ChillingworthT., DaviesR. M., FeltwellT.other authors2000; The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. Nature 403:665–668 [View Article][PubMed]
PeiZ., BlaserM. J.1993; PEB1, the major cell-binding factor of Campylobacter jejuni, is a homolog of the binding component in gram-negative nutrient transport systems. J Biol Chem 268:18717–18725[PubMed]
RoseA., KayE., WrenB. W., DallmanM. J.2012; The Campylobacter jejuni NCTC11168 capsule prevents excessive cytokine production by dendritic cells. Med Microbiol Immunol (Berl) 201:137–144 [View Article][PubMed]
SchultzJ. E., MatinA.1991; Molecular and functional characterization of a carbon starvation gene of Escherichia coli. J Mol Biol 218:129–140 [View Article][PubMed]
SchultzJ. E., LatterG. I., MatinA.1988; Differential regulation by cyclic AMP of starvation protein synthesis in Escherichia coli. J Bacteriol 170:3903–3909[PubMed]
ShawF. L., MulhollandF., Le GallG., PorcelliI., HartD. J., PearsonB. M., van VlietA. H. M.2012; Selenium-dependent biogenesis of formate dehydrogenase in Campylobacter jejuni is controlled by the fdhTU accessory genes. J Bacteriol 194:3814–3823 [View Article][PubMed]
StahlM., FriisL. M., NothaftH., LiuX., LiJ., SzymanskiC. M., StintziA.2011; L-fucose utilization provides Campylobacter jejuni with a competitive advantage. Proc Natl Acad Sci U S A 108:7194–7199 [View Article][PubMed]
VelayudhanJ., JonesM. A., BarrowP. A., KellyD. J.2004; L-serine catabolism via an oxygen-labile l-serine dehydratase is essential for colonization of the avian gut by Campylobacter jejuni.. Infect Immun 72:260–268 [View Article][PubMed]
WagenaarJ. A., Jacobs-ReitsmaW., HofshagenH., NewellD.2008; Poultry colonisation with Campylobacter and its control at the primary production level. In Campylobacter, 3rd edn. pp. 667–678 Edited by NachamkinI., SzymanskiC. M., BlaserM. J. Washington: American Society for Microbiology;
YenM.-R., TsengY.-H., SaierM. H.Jr2001; Maize Yellow Stripe1, an iron-phytosiderophore uptake transporter, is a member of the oligopeptide transporter (OPT) family. Microbiology 147:2881–2883[PubMed]
Campylobacter jejuni carbon starvation protein A (CstA) is involved in peptide utilization, motility and agglutination, and has a role in stimulation of dendritic cells