Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression data from Arabidopsis thaliana PHR1 primary targets genes under phosphate starvation stress

ABSTRACT: We performed a transcriptomic analysis of Pi starvation responses in Arabidopsis thaliana (Columbia-0) phr1 mutant plants expressing PHR1 in presence of cicloheximide, that inhibit protein translation, thus preventing any effect of PHR1 on the expression of indirect targets. Results show the primary target genes of PHR1 in the responses to Pi starvation. The analysis was performed in phr1 plants and phr1 plants overexpressing a fusion GR:PHR1 (OXGR:PHR1) whose activity is postranslationally controlled by dexamethasone (DEX), that allows studying gene expression upon PHR1 activation and the concomitant inhibition of translation with cycloheximide (CHX), thus preventing any effect of PHR1 on the expression of indirect targets. To perform this study, phr1 and OXGR:PHR1 phr1 plants were grown for seven days in complete Johnson liquid media (+Pi), 2 days in Pi-lacking media (-Pi) and then supplemented with 5 µM DEX and 10 µM CHX for 6 hours before harvesting. Three independent biological samples of total RNA from shoot and root were hybridized separately.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Regla Bustos

PROVIDER: E-GEOD-20955 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A central regulatory system largely controls transcriptional activation and repression responses to phosphate starvation in Arabidopsis.

Bustos Regla R Castrillo Gabriel G Linhares Francisco F Puga María Isabel MI Rubio Vicente V Pérez-Pérez Julian J Solano Roberto R Leyva Antonio A Paz-Ares Javier J

PLoS genetics 20100909 9

Plants respond to different stresses by inducing or repressing transcription of partially overlapping sets of genes. In Arabidopsis, the PHR1 transcription factor (TF) has an important role in the control of phosphate (Pi) starvation stress responses. Using transcriptomic analysis of Pi starvation in phr1, and phr1 phr1-like (phl1) mutants and in wild type plants, we show that PHR1 in conjunction with PHL1 controls most transcriptional activation and repression responses to phosphate starvation, ...[more]

PMID: 20838596

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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

Expression data from Arabidopsis thaliana PHR1 primary targets genes under phosphate starvation stress

Publications

A central regulatory system largely controls transcriptional activation and repression responses to phosphate starvation in Arabidopsis.

Similar Datasets

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