Transcriptional profiles of Campylobacter jejuni NCTC 11168 and JL272 to erythromycin treatment
Ontology highlight
ABSTRACT:
ORGANISM(S): Campylobacter jejuni
SUBMITTER: Qingqing Xia
PROVIDER: E-GEOD-43026 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Publications
Adaptive mechanisms of Campylobacter jejuni to erythromycin treatment.
Xia Qingqing Q Muraoka Wayne T WT Shen Zhangqi Z Sahin Orhan O Wang Hongning H Wu Zuowei Z Liu Peng P Zhang Qijing Q
BMC microbiology 20130614
<h4>Background</h4>Macrolide is the drug of choice to treat human campylobacteriosis, but Campylobacter resistance to this antibiotic is rising. The mechanisms employed by Campylobacter jejuni to adapt to erythromycin treatment remain unknown and are examined in this study. The transcriptomic response of C. jejuni NCTC 11168 to erythromycin (Ery) treatment was determined by competitive microarray hybridizations. Representative genes identified to be differentially expressed were further characte ...[more]
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Project description:Erythromycin is the drug of choice to treat campylobacteriosis, but resistance to this antibiotic is rising. The adaptive mechanisms employed by Campylobacter jejuni to erythromycin treatment remain unknown. The aim of this study is to determine the molecular basis underlying CampylobacterM-bM-^@M-^Ys immediate response to Ery treatment. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni wild type strain NCTC 11168. One hybridizations were performed: sham-treated NCTC 11168 v.s. lethal dose erythromycin treated NCTC 11168. Samples were independently grown and harvested. There were three biological replicates of each sample.
2013-10-10 | E-GEOD-43023 | biostudies-arrayexpress
Project description:Erythromycin is the drug of choice to treat campylobacteriosis, but resistance to this antibiotic is rising. The adaptive mechanisms employed by Campylobacter jejuni to erythromycin treatment remain unknown. The aim of this study is to determine the molecular basis underlying CampylobacterM-bM-^@M-^Ys immediate response to Ery treatment. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni wild type strain NCTC 11168. One hybridizations were performed: sham-treated NCTC 11168 v.s. sub-lethal dose erythromycin treated NCTC 11168. Samples were independently grown and harvested. There were three biological replicates of each sample.
2013-10-10 | E-GEOD-43024 | biostudies-arrayexpress
Project description:Erythromycin is the drug of choice to treat campylobacteriosis, but resistance to this antibiotic is rising. The adaptive mechanisms employed by Campylobacter jejuni to erythromycin treatment remain unknown. The aim of this study is to determine the molecular basis underlying CampylobacterM-bM-^@M-^Ys immediate response to Ery treatment. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni macrolide resistant strain JL272. One hybridizations were performed: sham-treated JL272 v.s. lethal dose erythromycin treated JL272. Samples were independently grown and harvested. There were three biological replicates of each sample.
2013-10-10 | E-GEOD-43025 | biostudies-arrayexpress
Project description:Campylobacter, a major foodborne pathogen, is increasingly resistant to macrolide antibibotics. Previous findings suggested that development of macrolide resistance in Campylobacter requires a multi-step process, but the molecular mechanisms involved in the process are not known. In our study, erythromycin-resistant C. jejuni mutant (R) was selected in vitro by stepwise exposure of C. jejuni NCTC11168(S) to increasing concentrations of erythromycin.The resistant were subjected to microarray and the the global transcriptional profile was analyzed. In this series, DNA microarray was used to compare the gene expression profiles of the macrolide-resistant strain with its parent wild-type strain NCTC11168. A large number of gene showed significant changes in R. The up-regulated genes in the resistant strains are involved in miscellaneous periplasmic proteins, efflux protein and putative aminotransferase, while the majority of the down-regulated genes are involved in electron transport, lipoprotein, heat shock protein and unknown function proteins. The over-expression of efflux pump and periplasmic protein was involved in the development of resistance to macrolide in C. jejuni. An eight chip study using total RNA recovered from four separate resistant-type cultures of Erythrocin-resistant Campylobacter jejuni NCTC111168 (R) and four separate cultures of Campylobacter jejuni NCTC111168 (S). Each chip measures the expression level of 1634 genes from Campylobacter jejuni NCTC11168.
2013-08-13 | E-GEOD-49257 | biostudies-arrayexpress
Project description:Cj0440c encodes a putative transcriptional regulator. To determine the role of Cj0440c in C.jejuni, we knocked out Cj0440c in the wild-type strain (S) to obtain the Cj0440c mutants (SM). Then we compared the transcriptome of the Cj0440c mutant with that of the parent strain using DNA microarray. These comparisons identified 19 genes that showed aM-bM-^IM-%2-fold change in expression in SM. The differentially expressed genes in SM encode proteins involved in flagellar biosynthesis, O-linked glycosylation and hypothetical proteins with unknown fuctions. Cj0440c may regulate flagellar structural element expression or as a compenent of flagellar complex co-expressed with other flagellar genes. Subsequent experiments demonstrated that inactivation of Cj0440c affected corresponding phenotypes of C.jejuni, including broken flagella, weaker motility and reduced colonization ability in chickens. These findings indicate that Cj0440c governs the expression of multiple genes related to flagellar biosynthesis and O-linked glycosylation. This study provides favorable evidence for completing the information of the Campylobacter jejuni genome. An eight chip study using total RNA recoverd from four separate wild-type cultures of Campylobacter jejuni NCTC111168 (S) and four separate cultures of a mutant strain, Campylobacter jejuni NCTC11168 delta- Cj0440c (SM), in which Cj0440c is deleted. Each chip measures the expression level of 1634 genes from Campylobacter jejuni NCTC11168.
2013-08-13 | E-GEOD-49256 | biostudies-arrayexpress
Project description:Cj0440c, a putative transcriptional regulator, was over-expressed in the high-level erythromycin-resistant (Eryr) Campylobacter jejuni strains. To determine the role of Cj0440c on the development and fitness of erythromycin resistance in C. jejuni, we knocked out Cj0440c in Eryr strain (R) to obtain the Cj0440c mutants (RM). Then we compared the transcriptome of the Cj0440c mutant with that of the parent strain using DNA microarray. These comparisons identified 9 genes that showed a M-bM-^IM-%2-fold change in expression in RM. The differentially expressed genes in RM are related to flagellar biosynthesis and unknown functions. What's more, katA, encoding catalase, down-regulated in RM. Cj0440c may progress flagellar genes expression, help to escape drug pressure and disseminate and colonize smoothly, and Cj0440c in Eryr Campylobacter may protect bacteria from harmful oxygen stress from the host immune system, other microorganism in host intestinal and its own products. These findings indicate that Cj0440c is essential for the fitness (growth) of resistant C. jejuni by controlling the expression of several genes involved in flagellar assembly and catalase, enhancing cell motility for colonization and invasion under the pressure of drug. This study widened our understanding on the molecular mechanism of resistance and provides scientific reference for drug research and application. An eight-chip study using total RNA recoverd from four separate resistant-type cultures of Erythrocin-resistant Campylobacter jejuni NCTC 111168 (R) and four separate cultures of a mutant strain, erythrocin-resistant Campylobacter jejuni NCTC 11168 delta- Cj0440c (RM), in which Cj0440c is deleted. Each chip measures the expression level of 1634 genes from Campylobacter jejuni NCTC 11168.
2013-08-13 | E-GEOD-49255 | biostudies-arrayexpress
Project description:During colonization in the host gastrointestinal tract, the enteric bacteria Campylobacter jejuni is exposed to a variety of signaling molecules including the catecholamine hormones epinephrine (Epi) and norepinephrine (NE). NE has been determined to stimulate the growth of C. jejuni as well as increase its pathogenicity. To investigate the mechanisms of NE or Epi on the biology of C. jejuni, the global gene expression profiles of C. jejuni NCTC 11168 cultured in iron-restricted medium were analyzed in response to NE or Epi. Totally, 183 and 156 genes were differentially expressed by NE and Epi respectively, with 102 differentially expressed genes common between the two treatments. These genes are involved in diverse cellular functions including iron uptake systems, motility, virulence, oxidative stress response, nitrosative stress tolerance, enzyme metabolism, DNA repair and metabolism and ribosomal protein biosynthesis. Adherence to and invasion of Caco-2 cells by C. jejuni were enhanced upon exposure to NE or Epi. These results indicated that NE and Epi have similar effects on the gene expression of C. jejuni and that the effects on gene expression may contribute to elucidate the mechanisms on interaction between host and C. jejuni. Transcriptional profiles were analyzed using microarray to compared the epinephrine (Epi) or norepinephrine (NE) treated and untreated Campylobacter jejuni NCTC 11168. NE or Epi treated and untreated cultures of C. jejuni NCTC 11168 were collected at the mid-log phase (?36 h cultures). Three (for NE or Epi treated culture) or four (for untreated control culture) independent biological replicates were performed. Total RNA was extracted using RiboPure™-Bacteria Kit (Ambion, Life Technologies) according to the manufacturer’s instructions. The quality and quantity of total RNA were determined by an Agilent Bioanalyzer 2100. Total RNA was amplified and labeled by Low Input Quick Amp Labeling Kit, One-Color, following the manufacturer’s instructions. Labeled cRNA were purified by RNeasy mini kit (QIAGEN, GmBH, Germany). Each Slide was hybridized with 600ng Cy3-labeled cRNA using Gene Expression Hybridization Kit (Agilent technologies, Santa Clara, CA, US) in Hybridization Oven, according to the manufacturer’s instructions. After 17 hours hybridization, slides were washed in staining dishes with Gene Expression Wash Buffer Kit (Agilent technologies, Santa Clara, CA, US), followed the manufacturer’s instructions. Slides were scanned by Agilent Microarray Scanner with default settings, Dye channel: Green, Scan resolution=5?m, PMT 100%, 10%, 16bit. Data were extracted with Feature Extraction software 10.7. Raw data were normalized by Quantile algorithm, Gene Spring Software 11.0. The genes with fold change ? 1.5 and P < 0.05 were selected as differentially expressed.
2015-02-10 | E-GEOD-65187 | biostudies-arrayexpress
Project description:Objectives: The aim of the study was to characterize (-)-α-Pinene, which is one of the chemical constituents of Alpinia katsumadai seed essential oil responsible for its resistance modulatory activity in Campylobacter jejuni. Methods: Broth microdilution method was used to evaluate the antimicrobial and resistance modulatory potential of (-)-α-Pinene and ethidium bromide accumulation assay to determine its efflux-inhibitory activity in subinhibitory concentration. The target efflux system was identified using knock-out mutants in several efflux related genes. Furthermore, the influence of subinhibitory concentration of (-)-α-Pinene on C. jejuni NCTC 11168 was investigated using microarray technology in order to elucidate the adaptive mechanism of bacteria to treatment with this phytochemical. Knock-out mutants of key adaptation genes were constructed and their role in adaptation to several stress factors, including (-)-α-Pinene, different osmolites and pH, was investigated using Biolog phenotypical microarrays and CFU counts. Results: (-)-α-Pinene was confirmed as highly efficient Campylobacter jejuni resistance modulator, due to its efflux inhibitory activity, which was significantly higher compared to reference inhibitors CCCP and reserpine. The CmeABC along with newly characterized CmeI (Cj1687) was confirmed as its main target efflux system. The transcriptional analysis indicated that the heat shock regulators HspR and HrcA are the main transcription regulators involved in adaptation to (-)-α-Pinene treatment. Conclusions: (-)-α-Pinene is a novel CmeABC and CmeI efflux inhibitor, which evokes the heat shock response in Campylobacter jejuni. Influence of (-)-α-Pinene on gene expression in C. jejuni NCTC 11168 was determined by expressional analysis and qRT-PCR. Exponential phase culture was adjusted to OD600=0.2 in MHB using spectrophotometer (Smart Spec, Bio-Rad, Hercules, CA, USA). Five ml of culture was treated with 62.5 mg/L of (-)-alpha-pinene dissolved in DMSO. Only DMSO (0.048 %) was added to untreated samples. Cultures were treated for 2 h, incubated microaerobically shaking (160 rpm) at 42°C. Experiments were carried out in 4 biological replicates. RNA Protect Bacteria reagent (Qiagen, Maryland, USA) was added to the culture and total RNA was isolated using RNeasy mini kit (Qiagen) and treated with Ambion® Turbo DNA-freeTM kit (Invitrogen, USA). Microarrays with 4751 probes targeting 1756 genes specific for Campylobacter jejuni subsp. jejuni NCTC 11168 (Mycroarray, Biodiscovery-LLC, MI, USA) were used for gene expression analysis. The cDNA was synthesized with random hexamers, SuperScriptTM III Reverse Transcriptase and amynoallyl dUTP (all supplied by Invitrogen, USA) and labeled with monofunctional NHS-ester dye Amersham C3 or Cy5 (GE Healthcare, Buckinghamshire, UK). Concentration of cDNA and labeling efficiency was determined spectrophotometrically with NanoDrop 1000. Four biological replicates were hybridized to four microarrays according to manufacturerâs protocol, incubated for 24 hours at 42â°C and scanned at 532-nm (Cy3) and 635-nm (Cy5) wavelengths using GenePix 4100A (Molecular Devices, Sunnyvale, CA) following the manufacturer's protocol. Fluorescence intensities of each spot were extracted using GenePix Pro 7.0 (Molecular Devices).
2015-11-19 | E-GEOD-59879 | biostudies-arrayexpress
Project description:We perfomed the transcriptomic analysis of differential genetic expression in the CosR-knockdown condition with a DNA microarray. CosR-specific antisense-PNA was used for CosR-knockdown. The results revealed that CosR significantly affected the expression of several genes involved in various cellular function in Campylobacter. Final goal of this study is to figure out the importance and the role of CosR in Campylobacter by identification of the CosR regulons. Two-condition experiment, WT vs. CosR-knockdown. Biological replicates: 3 wild type (control) replicates, 3 CosR-knockdown replicates. For preparing the total RNA, C. jejuni cells were grown at 42M-BM-0C in MH broth supplemented with 1.5 M-NM-<M CosR-PNA for CosR knockdown to the mid-exponential phase for approximately 8 hr with shaking.
2013-01-01 | E-GEOD-40201 | biostudies-arrayexpress
Project description:Strain specific growth of C. jejuni on fucose has been linked to a plasticity region of the chromosome (PR2) and confers a competitive advantage during intestinal colonization. Growth on fucose induces gene expression of PR2 genes, but the regulatory mechanism of the structural genes involved with fucose utilization is unknown. A mutant was constructed to examine the role of Cj0480c, a putative IclR-type transcriptional regulator, on PR2 gene expression. Transcriptional profiles of wild-type C. jejuni and the Cj0480c mutant strain grown without fucose may provide insight in to the extent of the fucose regulon and genes that are necessary for fucose utilization. The design utilized an available two color microarray slide for the entire transcriptome of Campylobacter jejuni wild type strain NCTC 11168. Each sample represents one competitive hybridization: wild-type NCTC 11168 v.s. Cj0480c isogenic mutant. There were four biological replicates of each sample with a dye swap introduced in alternating replicates. Samples were independently grown, treated and harvested.
2014-01-01 | E-GEOD-46808 | biostudies-arrayexpress