Dataset Information

Effects of Campylobacter jejuni CJIE1 prophage on protein expression experiment 2

ABSTRACT: Temperate bacteriophages (prophages) have recently been demonstrated in Campylobacter jejuni. However, what they do there is largely unknown. In the series of studies that are the subject of these submissions we have investigated the relative expression levels of proteins in C. jejuni isolates that differ in the presence or absence of the CJIE1 prophage. At the time of the initial investigations whole genome sequence data were not available for the isolates used, though DNA microarray data indicated that the isolates were very closely related. The overall project was carried out through four separate experiments. Previous work in the scientific literature indicated that growth on medium lacking blood but containing sodium deoxycholate induced the expression of at least some proteins associated with virulence and provided data thought to be of relevance to the virulence of the bacterium. The second set of experiments (experiment 2) therefore compared protein expression in 4-plex iTRAQ experiments using two isolates. Isolate 00-2425 carried the CJIE1 prophage while the second isolate, 00-2426, did not. Three replicate experiments were done. Each isolate was grown on Mueller Hinton agar base and Mueller Hinton agar containing 0.1% sodium deoxycholate.

OTHER RELATED OMICS DATASETS IN: PRJNA213842PRJNA213841PRJNA211797PRJNA214465

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Campylobacter Jejuni 10186

SUBMITTER: Clifford Clark

LAB HEAD: Garret R. Westmacott

PROVIDER: PXD000799 | Pride | 2014-07-31

REPOSITORIES: Pride

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Publications

The CJIE1 prophage of Campylobacter jejuni affects protein expression in growth media with and without bile salts.

Clark Clifford G CG Chong Patrick M PM McCorrister Stuart J SJ Simon Philippe P Walker Matthew M Lee David M DM Nguy Kimberly K Cheng Keding K Gilmour Matthew W MW Westmacott Garrett R GR

BMC microbiology 20140319

<h4>Background</h4>The presence of Campylobacter jejuni temperate bacteriophages has increasingly been associated with specific biological effects. It has recently been demonstrated that the presence of the prophage CJIE1 is associated with increased adherence and invasion of C. jejuni isolates in cell culture assays.<h4>Results</h4>Quantitative comparative proteomics experiments were undertaken using three closely related isolates with CJIE1 and one isolate without CJIE1 to determine whether th ...[more]

PMID: 24641125

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Project description:Temperate bacteriophages (prophages) have recently been demonstrated in Campylobacter jejuni. However, what they do there is largely unknown. In the series of studies that are the subject of these submissions we have investigated the relative expression levels of proteins in C. jejuni isolates that differ in the presence or absence of the CJIE1 prophage. At the time of the initial investigations whole genome sequence data were not available for the isolates used, though DNA microarray data indicated that the isolates were very closely related. The overall project was carried out through four separate experiments. The first set of experiments (experiment 1) therefore compared protein expression in 4-plex iTRAQ experiments using three isolates carrying the (previously sequenced) prophage with a fourth isolate that was epidemiologically and genetically related to these isolates but which naturally lacked the CJIE1 prophage. Three replicate experiments were done. These isolates were grown on medium containing blood to obtain optimal growth and determine whether any differences were found. Three replicate experiments were done. However, since these growth conditions may not mimic in vivo growth of the organism the results may not be considered relevant or very interesting. Previous work in the scientific literature indicated that growth on medium lacking blood but containing sodium deoxycholate induced the expression of at least some proteins associated with virulence and provided data thought to be of relevance to the virulence of the bacterium. Therefore two isolates were chosen, one with and one without the CJIE1 prophage. Each of three replicates of a 4-plex iTRAQ experiment compared protein expression of the two isolates (00-2425 and 00-2426) after growth on Mueller-Hinton agar with and without 0.1% sodium deoxycholate. There was pretty good overlap between the proteins expressed in the first two experiments but, because all results were expressed as log2 change values, it was difficult to determine whether the expression of some proteins was completely absent in one or the other isolate. As well, protein expression overall showed quite extensive differences compared with the first experiment that we attributed to the lack of blood in the growth medium; we thought we might be seeing a response to limited iron as well. Finally, the response to the deoxycholate was extensive, and we wished to further explore this phenomenon as a means of determining or verifying what effects were due to the presence of the CJIE1 prophage and which were due to other differences. Further 4-plex iTRAQ experiments compared protein expression for each of isolate after growth on medium containing blood, without blood, without blood plus sodium deoxycholate, and without blood plus 2.5% Oxgall. Comparison of the results for isolate 00-2425 (with the CJIE1 prophage) and 00-2426 (without the prophage) identified a small number of proteins that could not be detected in one or the other isolate and allowed discrimination of the effects of the prophage from the responses to iron limitation and bile salt preparations. This dataset is associated with the first experiment, experiment 1, which compares C. jejuni isolates carrying the CJIE1 prophage (00-2425, 00-2538, 00-2544) with a very closely related isolate lacking the prophage (00-2426)

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).

Dataset Information

Effects of Campylobacter jejuni CJIE1 prophage on protein expression experiment 2

Publications

The CJIE1 prophage of Campylobacter jejuni affects protein expression in growth media with and without bile salts.

Similar Datasets

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