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Genome-wide effect of cholera toxin and nitrate treatment in etiolated rice plants

ABSTRACT: The present study used microarray approach to identify the genomewide response to cholera toxin in the presence of nitrate. Considering that fact that the possibility of the existence of multiple Gα genes/proteins in plants has not been conclusively ruled out, analysis of the genomewide impact of RGA1 mutation in rice and GPA1 mutation in Arabidopsis reveal only those genes that are under their direct control. On the other hand, assuming that all those different Gα subunits in any given plant are regulated by cholera toxin, analysis of the genomwide response to cholera toxin could capture the entire G-protein responsive transcriptome, beyond what can be revealed by the mutant approach. This could reveal even those genes that respond to other, as yet unidentified Gα subunits, as well as reveal some genes that are non-specifically regulated by cholera toxin, independent of any G-proteins.

ORGANISM(S): Oryza sativa

PROVIDER: GSE20924 | GEO | 2010/12/01

SECONDARY ACCESSION(S): PRJNA124359

REPOSITORIES: GEO

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Genome-wide effect of cholera toxin and nitrate treatment in etiolated rice plants

Project description:The present study used microarray approach to identify the genomewide response to cholera toxin in the presence of nitrate. Considering that fact that the possibility of the existence of multiple GM-NM-1 genes/proteins in plants has not been conclusively ruled out, analysis of the genomewide impact of RGA1 mutation in rice and GPA1 mutation in Arabidopsis reveal only those genes that are under their direct control. On the other hand, assuming that all those different GM-NM-1 subunits in any given plant are regulated by cholera toxin, analysis of the genomwide response to cholera toxin could capture the entire G-protein responsive transcriptome, beyond what can be revealed by the mutant approach. This could reveal even those genes that respond to other, as yet unidentified GM-NM-1 subunits, as well as reveal some genes that are non-specifically regulated by cholera toxin, independent of any G-proteins. Total RNA was isolated from 10 day old excised rice leaves treated with or without cholera toxin in the presence of nitrate.Total RNA was first converted into cDNA and then into cRNA labeled with Cy3 and Cy5 dyes. These were hybridized on the 44K 60-mer whole genome arrays on glass slides. The slides were washed and scanned on an Agilent scanner (G2565B) at 100 % laser power. Data extraction was carried out with Agilent Feature Exiraction software (version 9.1). The experiment was repeated with a biological-cum-technical replicate where total RNA samples were isolated from a fresh batch of rice plants and the cRNAs were labeled with opposite dyes (dye-swap).

2010-12-01 | E-GEOD-20924 | biostudies-arrayexpress

Genetic analysis of the genome wide role of G-proteins in rice metabolism

Project description:The current study reports a genome-wide transcriptional profiling of the only known Gα mutants in rice (RGA1).

2010-12-01 | GSE20925 | GEO

co-immunoprecipitation of the Arabidopsis G protein β subunit AGB1

Project description:Heterotrimeric guanine nucleotide-binding (G) proteins are composed of Gα, Gβ, and Gγ subunits, and function as molecular switches in signal transduction. In Arabidopsis thaliana there are one canonical Gα (GPA1), three extra-large Gα (XLG1, XLG2 and XLG3), one Gβ (AGB1) and three Gγ (AGG1, AGG2 and AGG3) subunits. To elucidate AGB1 molecular signaling, we performed immunoprecipitation using plasma membrane enriched proteins followed by mass spectrometry to identify the protein interactors of AGB1. After eliminating proteins present in the control immunoprecipitation, commonly identified contaminants, and organellar proteins, a total of 103 candidate AGB1-associated proteins were confidently identified. We identified all of the G protein subunits except XLG1, receptor-like kinases (RLKs), Ca2+ signaling related proteins and 14-3-3-like proteins, all of which may couple with or modulate G protein signaling.

2018-10-02 | PXD011255 | Pride

Transcriptome profiling reveals a genomewide role of G-proteins

Project description:Knock out mutants of GPA1 (gpa1-5), GCR1 (gcr1-5) as well as their double mutant (gpa1-5 gcr1-5) were used to obtain genomewide transcriptome profile to deduce genomewide effect of both the subunits alone or in combination. Heterotrimeric G-protein signaling pathways have been implicated in a variety of biological processes in plants. While an impressive range of important plant responses are known to be mediated by G-protein signaling pathways, the genes that respond through these pathways are not yet known completely. More importantly, the role of G-proteins in mediating plant metabolic regulation, which is well established in animal systems, has not received adequate attention. The availability of loss-of-function G_alpha (GPA1) and G-protein coupled receptor (GCR1) mutant(s) as well as their double mutant in Arabidopsis provided an opportunity for a genome-wide microarray analysis in the current study for the first time.

2015-02-25 | GSE40217 | GEO

Transcriptome profiling reveals a genomewide role of G-proteins

2015-02-25 | E-GEOD-40217 | biostudies-arrayexpress

Transcriptome profiling of rice G-alpha mutant and WT in response to nitrate treatment

Project description:This study reports the genome-wide transcriptional profiling of the shoots of 1 month old Gα (RGA1) mutant and wild type rice plants grown in pots and treated with or without nitrate for 90 minutes after 2-days of N-free condition

2019-09-12 | GSE62164 | GEO

Total and cholera toxin B-tetramer specific T cells from Peyer's patches and mesenteric lymph nodes

Project description:T cells were separately isolated from MLN and PP from two naive mice, and T cells binding to a cholera toxin B subunit specific MHC II tetramer from mice orally immunized with cholera toxin 7 days or boosted 5 days with a second dose of cholera toxin. Cells were purified using flow cytometric cell sorting and 3´ (n=5) or 5´ (n=5) libraries were constructed using the 10X workflow. From 5´libraries TCR libraries were also constructed and sequenced.

2022-06-22 | E-MTAB-11805 | biostudies-arrayexpress

Transcriptional profile of differentially matured human monocyte-derived DCs

Project description:Human MoDCs were stimulated with TNF, Cholera Toxin or LPS to identify candidates for common genes expression.

2018-03-14 | GSE106080 | GEO

Host-derived O-glycans inhibit toxigenic conversion by a virulence-encoding phage in Vibrio cholerae

Project description:Pandemic and endemic strains of Vibrio cholerae arise from toxigenic conversion by the CTXφ bacteriophage, a process by which CTXφ infects non-toxigenic strains of V. cholerae. CTXφ encodes the cholera toxin, an enterotoxin responsible for the watery diarrhea associated with cholera infections. Despite the critical role of CTXφ during infections, signals that affect CTXφ-driven toxigenic conversion or expression of the CTXφ-encoded cholera toxin remain poorly characterized, particularly in the context of the gut mucosa. Here, we identify mucin polymers as potent regulators of CTXφ-driven pathogenicity in V. cholerae. Our results indicate that mucin-associated O-glycans block toxigenic conversion by CTXφ and suppress the expression of CTXφ-related virulence factors, including the toxin co-regulated pilus and cholera toxin, by interfering with the TcpP/ToxR/ToxT virulence pathway. By synthesizing individual mucin glycan structures de novo, we identify the Core 2 motif as the critical structure governing this virulence attenuation. Overall, our results highlight a novel mechanism by which mucins and their associated O-glycan structures affect CTXφ-mediated evolution and pathogenicity of V. cholerae, underscoring the potential regulatory power housed within mucus.

2022-12-01 | GSE213598 | GEO

Transcription profiling of pig IPEC-J2 cells treated with cholera toxin for 8h

Project description:The molecular mechanisms involved in host pathogen interactions at mucosal surfaces needs to be better understood in order to develop immune-mediated methods of protection against pathogens. Cholera toxin (CT) has the rare ability for a protein of inducing robust mucosal immunity in the gut and is therefore an excellent model with which to determine mechanisms of adjuvanticity and immunogenicity at intestinal mucosal surfaces. Jejunal epithelial cells are one of the first sites of antigen encounter. Therefore a porcine intestinal epithelial cell line, IPEC-J2, was cultured in 6-well transwell plates in the presence or absence of 50 ng/ml cholera toxin for up to 8 hours and the cell layer was harvested for gene expression analysis using the Affymetrix porcine genome array and real-time PCR analysis. Affymetrix analysis identified, and real-time PCR analysis of 15 genes confirmed, an increase in gene expression for 59 genes and a decrease in gene expression for 14 genes under CT treatment. An 8 hour time course of expression revealed that by 2-4 hours after CT treatment, all 10 upregulated genes were differentially expressed and by 4-6 hours after CT treatment 3 of the 5 downregulated genes were differentially expressed. These data suggest that the potent mucosal adjuvanticity and immunogenicity of CT derives from rapid alterations in gene expression at the site of first antigen encounter with the immune system. Characterization of early immune gene expression may elucidate potential biological mechanisms for mucosal immune induction leading to the development of effective vaccines against enteric pathogens. Experiment Overall Design: Confluent IPEC-J2 cell monolayers were treated with or without 50 ng/ml cholera toxin in transwell dishes for 8h. The experiment was repeated 3 times for a total of 6 chips, 3 cholera toxin treated and 3 untreated. Gene expression was compared between cholera toxin treated and untreated cells.

2008-06-11 | E-GEOD-10314 | biostudies-arrayexpress

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