ChIP-seq identified the regulation of genes by the quorum-sensing regulator OpaR in Vibrio parahaemolyticus
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ABSTRACT:
ORGANISM(S): Vibrio parahaemolyticus
PROVIDER: GSE122479 | GEO | 2020/10/27
REPOSITORIES: GEO
Similar Datasets
Project description:The quorum regulatory cascade is poorly characterized in Vibrio parahaemolyticus, in part because swarming and pathogenicity - the hallmark traits of the organism - are repressed by this scheme of gene control. As a consequence, many isolates appear silenced for quorum sensing via phase variation. In these studies, we examine a swarm proficient, virulent strain and find an altered function allele of the central quorum regulator luxO. We use this allele, which produces a constitutively active LuxO, to probe the upstream elements of the pathway and demonstrate their functionality for the first time. We find that the state of luxO affects expression of three small regulatory RNAS (Qrrs) and the activity of a translational fusion in opaR, the central output regulator. We use microarray profiling to determine the OpaR regulon, which was found to encompass ~5.2% of the genome. The quorum sensing proficient strain seems adapted for a sessile, community lifestyle; it is competent to uptake DNA, produces much capsular polysaccharide, has a high level of c-di-GMP, and strongly expresses one type six secretion system. Expressing the entire surface sensing regulon and numerous methyl accepting chemotaxis proteins, the quorum-disrupted cell type seems prepared for a mobile lifestyle. It is also cytotoxic to host cells in co-culture and expresses distinct type six as well as type three secretion systems. Thus, the scope and nature of the genes in the OpaR regulon provide many clues to the distinguishing traits of this Vibrio species as well as to the quite divergent survival strategies of the quorum ON/OFF phase variants The gene expression profiles of different strains of Vibrio parahaemolyticus cells grown on rich medium and compared using Affymetrix custom microarrays.
2011-12-01 | E-GEOD-28216 | biostudies-arrayexpress
Project description:The quorum regulatory cascade is poorly characterized in Vibrio parahaemolyticus, in part because swarming and pathogenicity - the hallmark traits of the organism - are repressed by this scheme of gene control. As a consequence, many isolates appear silenced for quorum sensing via phase variation. In these studies, we examine a swarm proficient, virulent strain and find an altered function allele of the central quorum regulator luxO. We use this allele, which produces a constitutively active LuxO, to probe the upstream elements of the pathway and demonstrate their functionality for the first time. We find that the state of luxO affects expression of three small regulatory RNAS (Qrrs) and the activity of a translational fusion in opaR, the central output regulator. We use microarray profiling to determine the OpaR regulon, which was found to encompass ~5.2% of the genome. The quorum sensing proficient strain seems adapted for a sessile, community lifestyle; it is competent to uptake DNA, produces much capsular polysaccharide, has a high level of c-di-GMP, and strongly expresses one type six secretion system. Expressing the entire surface sensing regulon and numerous methyl accepting chemotaxis proteins, the quorum-disrupted cell type seems prepared for a mobile lifestyle. It is also cytotoxic to host cells in co-culture and expresses distinct type six as well as type three secretion systems. Thus, the scope and nature of the genes in the OpaR regulon provide many clues to the distinguishing traits of this Vibrio species as well as to the quite divergent survival strategies of the quorum ON/OFF phase variants
2011-12-01 | GSE28216 | GEO
Project description:Vibrio parahaemolyticus an emerging pathogen that is a causative agent of foodborne gastroenteritis when raw or undercooked seafood is consumed. Previous microarray data using a Vibrio parahaemolyticus RIMD2210633 chip has shown the master quorum-sensing regulator OpaR controls virulence, type III and type VI secretion systems, and flagellar and capsule production genes. In a parallel study, RNA-Seq was used to comparatively study the transcriptome changes of wild type Vibrio parahaemolyticus BB22 and a ΔopaR strain directly. Differences in mRNA expression were analyzed using next generation Illumina sequencing and bioinformatics techniques to align and count reads. A comparison with the previous microarray data showed good correlation between the shared genes. The RNA-Seq offered an insight into control of genes specific to the Vibrio parahaemolyticus BB22 strain as well as a new look at the sRNAs that are expressed. Eleven transcriptional regulators with greater than 4 fold regulation in the previous microarray study and 2 fold regulation in the RNA-Seq analysis, were chosen to validate the data using qRT-PCR and further characterized with electrophoretic mobility shift assays (EMSAs) to determine if they are direct targets of OpaR. The transcription factors chosen play key roles in virulence, surface motility, cell to cell interactions, and cell surface characteristics. One small RNA was identified in the RNA-Seq data to be quorum-sensing controlled and unidentified by other programs. The RNA-Seq data has aided in understanding and elucidating the hierarchy of quorum-sensing control of OpaR in Vibrio parahaemolyticus. The wild type Vibrio parahaemolyticus BB22 strain LM5312 and an opaR deletion strain LM5674 were analyzed for mRNA expression using RNA-Seq.
2013-12-31 | E-GEOD-53639 | biostudies-arrayexpress
Project description:Vibrio parahaemolyticus an emerging pathogen that is a causative agent of foodborne gastroenteritis when raw or undercooked seafood is consumed. Previous microarray data using a Vibrio parahaemolyticus RIMD2210633 chip has shown the master quorum-sensing regulator OpaR controls virulence, type III and type VI secretion systems, and flagellar and capsule production genes. In a parallel study, RNA-Seq was used to comparatively study the transcriptome changes of wild type Vibrio parahaemolyticus BB22 and a ΔopaR strain directly. Differences in mRNA expression were analyzed using next generation Illumina sequencing and bioinformatics techniques to align and count reads. A comparison with the previous microarray data showed good correlation between the shared genes. The RNA-Seq offered an insight into control of genes specific to the Vibrio parahaemolyticus BB22 strain as well as a new look at the sRNAs that are expressed. Eleven transcriptional regulators with greater than 4 fold regulation in the previous microarray study and 2 fold regulation in the RNA-Seq analysis, were chosen to validate the data using qRT-PCR and further characterized with electrophoretic mobility shift assays (EMSAs) to determine if they are direct targets of OpaR. The transcription factors chosen play key roles in virulence, surface motility, cell to cell interactions, and cell surface characteristics. One small RNA was identified in the RNA-Seq data to be quorum-sensing controlled and unidentified by other programs. The RNA-Seq data has aided in understanding and elucidating the hierarchy of quorum-sensing control of OpaR in Vibrio parahaemolyticus.
2013-12-31 | GSE53639 | GEO
Project description:Vibrio parahaemolyticus is the causative agent of food-borne gastroenteritis disease. Once consumed, human acid gastric fluid is perhaps one of the most important environmental stresses imposed on the bacterium. Herein, for the first time, we investigated Vibrio parahaemolyticus CHN25 response to artificial gastric fluid (AGF) stress by transcriptomic analysis. The bacterium at logarithmic growth phase (LGP) displayed lower survival rates than that at stationary growth phase (SGP) under a sub-lethal acid condition (pH 4.9). Transcriptome data revealed that 11.6% of the expressed genes in Vibrio parahaemolyticus CHN25 was up-regulated in LGP cells after exposed to AGF (pH 4.9) for 30 min, including those involved in sugar transport, nitrogen metabolism, energy production and protein biosynthesis, whereas 14.0% of the genes was down-regulated, such as ABC transporter and flagellar biosynthesis genes. In contrast, the AGF stress only elicited 3.4% of the genes from SGP cells, the majority of which were attenuated in expression. Moreover, the number of expressed regulator genes was also substantially reduced in SGP cells. Comparison of transcriptome profiles further revealed forty-one growth-phase independent genes in the AGF stress, however, half of which displayed distinct expression features between the two growth phases. Vibrio parahaemolyticus seemed to have evolved a number of molecular strategies for coping with the acid stress. The data here will facilitate future studies for environmental stresses and pathogenicity of the leading food-borne pathogen worldwide. When V.parahemolyticus CHN25 grown to log phase and stationary phase at 37°C in TSB-3% NaCl, different cultures were subsequently exposed to artificial gastric fluid at 37°C for 30 min. Two independent experiments were performed at each phase for microarray expreriments.
2015-06-30 | E-GEOD-63167 | biostudies-arrayexpress
Project description:Naturally bacteria are commonly forced to remain in stationary phase. There is no increase in cell mass, however, cell division keep on. Vibrio (V.) parahaemolyticus is an aquatic bacterium capable of causing foodborne gastroenteritis outbreaks all over the world. So far, little is known about whole genomic expression of V. parahaemolyticus in the early stationary phase compared with the phase of exponential growth. Since under starvation cell sizes decrease and endogenous metabolism reduces, genes are considered to be highly repressed in the stationary phase. However, our data shows in total 172 induced genes, while 61 genes were repressed in the early stationary phase compared with exponential phase. In fatty acid and phospholipid metabolism functional category only induced genes were found, whereas in three other metabolic functional groups appeared no significant up-regulated genes (adjusted P-value<0.05). Genes in two metabolic functional categories remained stable in the early stationary phase. DAVID analyses were carried out exploring the gene regulation. In total, ten functional categories showed a total up-regulation in early stationary phase, while only three metabolic functional categories showed a down-regulation and four categories showed stably in early stationary phase.
2015-01-31 | GSE65448 | GEO
Project description:Naturally bacteria are commonly forced to remain in stationary phase. There is no increase in cell mass, however, cell division keep on. Vibrio (V.) parahaemolyticus is an aquatic bacterium capable of causing foodborne gastroenteritis outbreaks all over the world. So far, little is known about whole genomic expression of V. parahaemolyticus in the early stationary phase compared with the phase of exponential growth. Since under starvation cell sizes decrease and endogenous metabolism reduces, genes are considered to be highly repressed in the stationary phase. However, our data shows in total 172 induced genes, while 61 genes were repressed in the early stationary phase compared with exponential phase. In fatty acid and phospholipid metabolism functional category only induced genes were found, whereas in three other metabolic functional groups appeared no significant up-regulated genes (adjusted P-value<0.05). Genes in two metabolic functional categories remained stable in the early stationary phase. DAVID analyses were carried out exploring the gene regulation. In total, ten functional categories showed a total up-regulation in early stationary phase, while only three metabolic functional categories showed a down-regulation and four categories showed stably in early stationary phase. Early stationary phase gene expression was detected in total bacterial RNA of V. parahaemolyticus. Two phases (exponential phase and early stationary phase) were used in 8 biological replicates. Gene expression in exponential phase was used for normalization.
2015-01-31 | E-GEOD-65448 | biostudies-arrayexpress
Project description:Vibrio parahaemolyticus is the leading bacterial cause for seafood-related gastroenteritis worldwide. As an intestinal pathogen, V. parahaemolyticus competes with other commensal bacteria for the same pool of nutrients. The major source of nutrition for intestinal bacteria is intestinal mucus. We wanted to determine the expression profile of wild-type V. parahaemolyticus in mouse intestinal mucus and then perform a differential expression analysis in a ∆luxO deletion mutant, in which the high cell density quorum sensing regulator OpaR is constitutively expressed and low cell density regulator AphA is repressed.
2017-01-13 | GSE93547 | GEO
Project description:Comparative proteomics to identify proteins found in the media of Vibrio parahaemolyticus RIMD 2210633 bacteria with an active T6SS2 compared to bacteria with inactive T6SS2. Bacteria with an active T6SS2 are Vibrio parahaemolyticus RIMD 2210633 inwhich hcp1 was deleted to inactivate T6SS1. T6SS2 inactive bacteria are the former strain with an additional deletion in hcp2. Both strains express TfoX from an arabinose-inducible plasmid to induce T6SS2 activity.
2023-02-05 | PXD037980 | Pride
Project description:Compare the secreted proteins of a wild-type Vibrio parahaemolyticus strain with those of a mutant in hcp2, rendering the T6SS2 inactive
2023-02-05 | PXD037864 | Pride