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Campylobacter jejuni Cj1000 regulon

ABSTRACT: Transcriptional regulation mediates adaptation of pathogens to environmental stimuli and is important for host colonisation. The Campylobacter jejuni genome sequence reveals a surprisingly small set of regulators, mostly of unknown function, suggesting an intricate regulatory network. Interestingly, C. jejuni lacks the homologues of ubiquitous regulators involved in stress response found in many other Gram-negative bacteria. Nonetheless, cj1000 is predicted to code for the sole LysR-type regulator in the C. jejuni genome, and thus may be involved in major adaptation pathways. A cj1000 mutant strain was constructed and found to be attenuated in its ability to colonise 1-day old chicks. Complementation of cj1000 mutation restored the colonisation ability to that of wild type levels. The mutant strain was also outcompeted in a competitive colonisation assay of the piglet intestine. High resolution oxygraphy was carried out for the first time on C. jejuni and revealed a role for Cj1000 in controlling O2 consumption. Furthermore, microarray analysis of the cj1000 mutant revealed both direct and indirect regulatory targets, including genes involved in energy metabolism and oxidative stress defences. These results highlight the importance of Cj1000 regulation in host colonisation and in major physiological pathways.

ORGANISM(S): Campylobacter jejuni

PROVIDER: GSE43792 | GEO | 2013/07/03

SECONDARY ACCESSION(S): PRJNA187449

REPOSITORIES: GEO

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Campylobacter jejuni Cj1000 regulon

Project description:Transcriptional regulation mediates adaptation of pathogens to environmental stimuli and is important for host colonisation. The Campylobacter jejuni genome sequence reveals a surprisingly small set of regulators, mostly of unknown function, suggesting an intricate regulatory network. Interestingly, C. jejuni lacks the homologues of ubiquitous regulators involved in stress response found in many other Gram-negative bacteria. Nonetheless, cj1000 is predicted to code for the sole LysR-type regulator in the C. jejuni genome, and thus may be involved in major adaptation pathways. A cj1000 mutant strain was constructed and found to be attenuated in its ability to colonise 1-day old chicks. Complementation of cj1000 mutation restored the colonisation ability to that of wild type levels. The mutant strain was also outcompeted in a competitive colonisation assay of the piglet intestine. High resolution oxygraphy was carried out for the first time on C. jejuni and revealed a role for Cj1000 in controlling O2 consumption. Furthermore, microarray analysis of the cj1000 mutant revealed both direct and indirect regulatory targets, including genes involved in energy metabolism and oxidative stress defences. These results highlight the importance of Cj1000 regulation in host colonisation and in major physiological pathways. Microarray data was collected from three independent biological replicates and 3-9 technical replicates for each biological replicate.

2013-07-03 | E-GEOD-43792 | biostudies-arrayexpress

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Project description:C. jejuni HPC5 is a Campylobacter strain isolated from chickens. Following bacteriophage CP34 treatment on chickens colonised by C. jejuni HPC5, a series of CP34 insensitive strains like C. jejuni HPC5 R14 and C. jejuni HPC5 R20 were obtained which compromised their ability to colonise chickens. Reintroduction of C. jejuni HPC5 R14 and R20 in to chickens led to reversion of these strains and the MRPs of the revertant strains fell in to different classes termed C. jejuni HPC5 R14A, R14B, R20A, R20B and R20C and these strained were tested positive for colonisation proficient and bacteriophage sensitive. There are three biological replicates for each experiment. Seven independent cDNA preps of C. jejuni HPC5 were prepared and they were labelled independently using AF555. The labelled cDNAs were mixed together and this acted as the control. Each daughter strains were considered as sample and they were labelled with AF645. Hybridisations were done between C. jejuni HPC5 and C. jejuni HPC5 as one of the sample and each of the daughter strains in triplicates. The supplementary files (linked at the foot of this record) contain the summary of the replicate data of the corresponding strain (3 replicates/strain) which have been averaged.

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Relating differences in the colonisation potential of verotoxigenic Escherichia coli (VTEC) for plant hosts to wholescale genes in gene expression

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Relating differences in the colonisation potential of verotoxigenic Escherichia coli (VTEC) for plant hosts to wholescale genes in gene expression (2)

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Project description:In Campylobacter jejuni CmeR functions as a transcriptional repressor modulating the expression of the multidrug efflux pump CmeABC, which plays an important role in the resistance to antimicrobial agents and bile compounds. Using DNA microarray, we identified multiple genes that are either activated or repressed by CmeR in C. jejuni. The DNA microarray data was independently confirmed by quantitative real-time RT-PCR. The CmeR-regulated genes encode products of diverse functions including membrane proteins, drug efflux transporters, the C4-dicarboxylate transport/utilization system, and enzymes involved in the biosynthesis of capsular polysaccharide (CPS). Immunoblotting and Alcian blue staining further showed that CPS production is reduced in the cmeR mutant, confirming the regulation of CPS production by CmeR. Electrophoretic mobility shift assay revealed that recombinant CmeR bound specifically to several intergenic regions in the CPS gene cluster, suggesting that CmeR directly regulates this gene cluster. In the chicken host, the mutant carrying a null mutation in cmeR was severely outcompeted by the isogenic wild-type strain. Together these data indicate that CmeR functions as a global regulator in C. jejuni, modulates the expression of genes encoding diverse functions, and is important for the fitness of Campylobacter in the intestinal tract. Keywords: cell type comparison

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