Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Exploring signal-induced cellular regulatory subnetworks by use of partial least square regression (PLSR) multidimensional analysis of gene expression time series data

ABSTRACT: By use of time series microarray experiment we identified the characteristic temporal behaviour of ~2000 new gastrin-regulated genes (accession numbers: GSE32869). To inform gastrin regulatory networks with an additional experimental dimension, we determined the temporal transcriptome in the presence of the protein synthesis inhibitor cycloheximide (CHX). For those genes where the gastrin response in the presence of CHX was higher than the response with CHX alone the gene was classified as a primary gene not dependent on de novo protein synthesis for gastrin-induced expression.Gastrin induced genes that were not upregulated in the presence of CHX were classified as secondary genes, i.e., genes whose transcriptional activation dependends on de novo protein synthesis of one or several factors that must be induced by gastrin previous to these secondary genes.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Arnar Flatberg

PROVIDER: E-MTAB-1268 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The duration of gastrin treatment affects global gene expression and molecular responses involved in ER stress and anti-apoptosis.

Selvik Linn-Karina M LK Fjeldbo Christina S CS Flatberg Arnar A Steigedal Tonje S TS Misund Kristine K Anderssen Endre E Doseth Berit B Langaas Mette M Tripathi Sushil S Beisvag Vidar V Lægreid Astrid A Thommesen Liv L Bruland Torunn T

BMC genomics 20130628

<h4>Background</h4>How cells decipher the duration of an external signal into different transcriptional outcomes is poorly understood. The hormone gastrin can promote a variety of cellular responses including proliferation, differentiation, migration and anti-apoptosis. While gastrin in normal concentrations has important physiological functions in the gastrointestine, prolonged high levels of gastrin (hypergastrinemia) is related to pathophysiological processes.<h4>Results</h4>We have used geno ...[more]

PMID: 23805861

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Project description:A glucocorticoid-regulated BBM protein (35S:BBM-GR) was used in combination with microarray analysis to identify genes directly activated by BBM. We employed the system described by (Lloyd et al., 1994) in which dexamethasone (DEX) and cycloheximide (CHX) are applied together to respectively, induce nuclear localization of the BBM-GR protein and prevent translation of the primary targets mRNAs. In this way it is possible to identify direct targets of a transcriptional activator by comparing gene expression profiles between DEX+CHX-treated transgenic and wild-type tissues. The ability of the 35S:BBM GR construct to induce somatic embryogenesis in Arabidopsis seedlings was determined by phenotypic observation of 35S:BBM GR seeds germinated and grown in the presence of 10 ÂµM dexamethasone (DEX). As in 35S:BBM seedlings, we observed somatic embryo formation on the cotyledons, first leaves and shoot meristem of DEX-treated 35S:BBM GR seedlings. We identified a set of 20 genes (including BBM itself) and our analysis indicates that BBM directly activates a signaling pathway comprising transcription factors and other signaling molecules, but which does not initially include genes known to induce somatic embryogenesis, such as LEC1, LEC2 or WUS. The functions of the BBM target genes are unknown, however a number of them have recently been identified in microarray screens for meristem-expressed genes. The identification of BBM-interacting partners and downstream targets provides new tools for unraveling pathways related to plant cell growth and organogenesis. Keywords: transcriptional activation To identify candidate BBM targets, we treated 4 day old 35S:BBM-GR seedlings for 8 hours with 10 ÂµM DEX in the presence of 10 ÂµM CHX and compared the mRNA population from these seedlings using Arabidopsis Operon microarrays (http://www.ag.arizona.edu/microarray/) to that of 4 day old wild-type seedlings treated in the same way. The seed batches used in this analysis showed 100% EFS when grown continuously on media with 10 ÂµM DEX. Four day old seedlings were chosen as the experimental material because they are highly responsive to BBM GR activation and provide sufficient RNA for the microarray experiments. The biological reproducibility of the data was monitored using two independent single locus 35S:BBM GR lines. The technical reproducibility of the data was monitored using two independently DEX + CHX-treated samples from each transgenic line. Dye effects were monitored in a dye-swap experiment using one of the four RNA samples from the two biological replicates

Dataset Information

Exploring signal-induced cellular regulatory subnetworks by use of partial least square regression (PLSR) multidimensional analysis of gene expression time series data

Publications

The duration of gastrin treatment affects global gene expression and molecular responses involved in ER stress and anti-apoptosis.

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