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Calcium is a global regulator in Vibrio parahaemolyticus

ABSTRACT: Here we probe the global response to calcium in the marine bacterium and pathogen Vibrio parahaemolyticus by using transcriptome, reporter and phenotypic analyses. Swarming gene expression and motility were enhanced by calcium. Calcium also stimulated expression and function of one of the organism’s two type three secretion systems (T3SS1 but not the T3SS2). Although low calcium is an inducing signal for the T3SS of many organisms, calcium stimulation of T3SS has not been reported before. EGTA was also a stimulus for T3SS1 expression and activity; however this appeared to be the consequence of iron rather than calcium chelation. LafK, a key regulator of swarming genes, was found necessary for calcium induction of T3SS1 gene transcription and cytotoxicity. Regulation of swarming and T3SS1 were additionally linked by a negative feedback loop propagated by ExsA. Overexpression of exsA was used to probe the extent of the T3SS1 regulon and verify its coincident induction by calcium and EGTA. The calcium transcriptome analysis revealed a calcium-repressed LysR-type transcriptional regulator; CalR was shown to repress swarming and T3SS1 gene expression. Thus in V. parahaemolyticus, calcium influences gene expression and behavior and seems a signal pertinent for surface colonization and virulence.

ORGANISM(S): Vibrio parahaemolyticus RIMD 2210633 Vibrio parahaemolyticus

PROVIDER: GSE22190 | GEO | 2010/11/05

SECONDARY ACCESSION(S): PRJNA127551

REPOSITORIES: GEO

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Calcium is a global regulator in Vibrio parahaemolyticus

Project description:Here we probe the global response to calcium in the marine bacterium and pathogen Vibrio parahaemolyticus by using transcriptome, reporter and phenotypic analyses. Swarming gene expression and motility were enhanced by calcium. Calcium also stimulated expression and function of one of the organismM-bM-^@M-^Ys two type three secretion systems (T3SS1 but not the T3SS2). Although low calcium is an inducing signal for the T3SS of many organisms, calcium stimulation of T3SS has not been reported before. EGTA was also a stimulus for T3SS1 expression and activity; however this appeared to be the consequence of iron rather than calcium chelation. LafK, a key regulator of swarming genes, was found necessary for calcium induction of T3SS1 gene transcription and cytotoxicity. Regulation of swarming and T3SS1 were additionally linked by a negative feedback loop propagated by ExsA. Overexpression of exsA was used to probe the extent of the T3SS1 regulon and verify its coincident induction by calcium and EGTA. The calcium transcriptome analysis revealed a calcium-repressed LysR-type transcriptional regulator; CalR was shown to repress swarming and T3SS1 gene expression. Thus in V. parahaemolyticus, calcium influences gene expression and behavior and seems a signal pertinent for surface colonization and virulence. The gene expression profiles of Vibrio parahaemolyticus cells grown on rich medium or medium supplemented with CaCl2 or EGTA were compared using Affymetrix custom microarrays.

2010-11-05 | E-GEOD-22190 | biostudies-arrayexpress

Transcriptome Analysis of T3SS1 in Vibrio parahaemolyticus

Project description:Comparative transcriptional mRNA profiles were generated of bacterial pathogen Vibrio parahaemolyticus under conditions that induce activity of virulence factor type III secretion system 1 (T3SS1). Induction conditions included growth of the bacteria in Dulbecco's Modified Eagle Medium (DMEM), overexpression of transcriptional activator exsA and contact with HeLa cells in Hank's Balanced Salt Solution (HBSS), while non-inducing conditions included growth in Luria-Bertani medium supplemented with 2.5% (w/v) NaCl (LB-S) and overexpression of transcriptional anti-activator exsD. Transcriptome profiles of induction conditions were cross-compared against background non-inducing conditions and time points during HeLa cell infection (2 hr, 3 hr, 4 hr, 6 hr, 8 hr) were compared against pre-infection (0 hr) to identify genes important in T3SS1 activity and pathogenesis. Duplicate mRNA transcriptome profiles of V. parahaemolyticus grown in T3SS1 inducing (DMEM, exsA expression) and non-inducing (LB-S, exsD expression) conditions as well as during HeLa cell infection (0 hr, 2 hr, 3 hr, 4 hr, 6 hr, 8 hr) were generated by deep sequencing using an Illumina MiSeq

2014-01-21 | E-GEOD-51423 | biostudies-arrayexpress

Transcriptome Analysis of T3SS1 in Vibrio parahaemolyticus

2014-01-21 | GSE51423 | GEO

RNA-seq for PA4595

Project description:Pseudomonas aeruginosa is an important human pathogen which uses the type III secretion system 21 (T3SS) as a primary virulence factor to establish infections in humans. The results presented in this 22 report revealed that the ATP-binding protein PA4595 (named ArtR, a Regulator that is an ATP-activated 23 Repressor of T3SS) represses T3SS expression in P. aeruginosa. The expression of T3SS genes, 24 including exoS, exoY, exoT, exsCEBA and exsD-pscB-L, increased significantly when artR was 25 knockout. The effect of ArtR on ExsA is at the transcriptional level, not at the translational level. The 26 regulatory role and cytoplasm localization of ArtR suggest it belongs to the REG sub-family of 27 ATP-binding cassette (ABC) family. Purified GST-tagged ArtR showed ATPase activity in vitro. The 28 conserved aspartate residues in the dual Walker B motifs prove to be essential for the regulatory 29 function of ArtR. The regulation of T3SS by ArtR is unique, which does not involve the known 30 GacS/A-RsmY/Z-RsmA-ExsA pathway or Vfr. This is the first REG subfamily of ATP-binding 31 cassette that is reported to regulate T3SS genes in bacteria. The results specify a novel player in the 32 regulatory networks of T3SS in P. aeruginosa.

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Cytotoxic Vibrio T3SS1 Rewires Host Gene Expression to Subvert Cell Death Signaling and Activate Cell Survival Networks

Project description:Abstract: The type three secretion system 1 (T3SS1) of the marine bacterium, Vibrio parahaemolyticus, orchestrates a host cell death comprised of distinct stages defined by the action of specific effectors. To understand how the host responds to the T3SS1, we analyzed gene expression changes over time in infected primary fibroblasts by RNA-seq. The host cell’s transcriptional response was rapid, robust and temporally distinct. Despite the lethal action of the T3SS1 effectors, network analysis indicated that the integrated host transcriptional response to T3SS1 effectors activated cell survival and repressed cell death networks, suggesting that T3SS1 masks its cytotoxicity to the host cell. Pathway analysis revealed that signaling through several pathways, including MAPK pathways, was specifically altered by T3SS1. Overall, this temporal analysis of T3SS1-mediated subversion of host cell signaling revealed cytoprotective pathways that are likely hijacked by many bacterial pathogens to ensure survival. Methods: We employed genome-wide transcriptional profiling methods (RNA-seq) to measure changes in gene expression over time in response to infection with POR4:T3SS1- or POR3:T3SS1+. Time points were collected 45, 60, 75 and 90 minutes post-infection. In total, libraries from 27 samples (triplicates of uninfected, POR3/4 at t=45, 60, 75, and 90 minutes) were sequenced and mapped to the human genome. EdgeR was used for differential expression analysis using statistical cutoffs of of false discovery rate (FDR) ≤0.01, log2counts-per-million (log2CPM) ≥0 and fold change (FC) cutoffs of -1.5≥ FC ≥1.5

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Effect of calcium depletion on cold induced gene expression in barley

Project description:Agilent 44k barley microarray was used to screen calcium dependent cold induced genes. Calcium chelator (EGTA) or calcium channel blocker (lanthanum) or phospholipase C activator (mastoparan) pretreatments were performed to block/enhance calcium response in barley seedlings. Calcium depleted and control plants were cold treated and the difference in the gene expression was investigated.

2014-12-31 | GSE47432 | GEO

Vibrio parahaemolyticus

Project description:Transcriptome Analysis of T3SS1 in Vibrio parahaemolyticus

| PRJNA223069 | ENA

extracellular calcium chelation experiment_P. falciparum (lab strain 3d7) schizonts untreated vs P. falciparum (lab strain 3d7) reference RNA pool

Project description:Calcium is a universal second messenger molecule which plays a significant role in several biological processes. Presence of calcium sensors (calmodulins) and calcium-dependent protein kinases in Plasmodium species suggests an important role of calcium-dependent signaling pathways in the regulation of cellular processes in the malaria parasites. Evidence for extracellular calcium chelation of the control Plasmodium falciparum asexual blood stages not treated with the calcium ionophore ionomycin and EGTA has been presented here.

2012-07-15 | GSE33834 | GEO

extracellular calcium chelation experiment_P. falciparum (lab strain 3d7) schizonts treated with EGTA and ionomycin vs P. falciparum (lab strain 3d7) reference RNA pool

Project description:Calcium is a universal second messenger molecule which plays a significant role in several biological processes. Presence of calcium sensors (calmodulins) and calcium-dependent protein kinases in Plasmodium species suggests an important role of calcium-dependent signaling pathways in the regulation of cellular processes in the malaria parasites. Evidence for extracellular calcium chelation of the control Plasmodium falciparum asexual blood stages treated with the calcium ionophore EGTA ans later with ionomycin has been presented here.

2012-07-15 | GSE33836 | GEO

Surface sensing in Vibrio parahaemolyticus triggers a programme of gene expression that promotes colonization and virulence

Project description:Vibrio parahaemolyticus senses surfaces via impeded rotation of its polar flagellum. We have exploited this surface-sensing mechanism to trick the organism into thinking it is on a surface when it is growing in liquid. This facilitated studies of global gene expression in a way that avoided many of the complications of surface-to-liquid comparisons, and illuminated ~ 70 genes that respond to surface sensing per se. Almost all are surface-induced (not repressed) and encode swarming motility proteins, virulence factors or sensory enzymes involved with chemoreception and c-di-GMP signalling. Follow-up studies were performed to place the surfaceresponsive genes in a regulatory hierarchy. Mapping the hierarchy revealed two surprises about LafK, a transcriptional activator that until now has been considered to be the master regulator for the lateral flagellar system. First, LafK controls a more diverse set of genes than previously appreciated. Second, some laf genes are not under LafK control, which means LafK is not the master regulator after all. Additional experiments motivated by the transcriptome analyses revealed that growth on a surface lowers c-di-GMP levels and enhances cytotoxicity. Thus, we demonstrate that V. parahaemolyticus can invoke a programme of gene control upon encountering a surface and the specific identities of the surfaceresponsive genes are pertinent to colonization and pathogenesis. The gene expression profiles of swarming and swimming cell types of Vibrio parahaemolyticus were compared using Affymetrix custom microarrays.

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