Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Gene expression changes in response to the overexpression of the Arabidopsis RKD4 gene

ABSTRACT: The Arabidopsis RWP-RK protein RKD4 is expressed specifically in early embryogenesis and triggers embryonic cell division sequences. We used Affymetrix ATH1 microarrays to analyze the pattern of gene expression changes in response to induced ectopic expression of RKD4 in post-embryonic organs. Plants of a dexamethasone (DEX)-inducible overexpression line and a control line were grown under normal conditions for 2 days and then treated with DEX for 1 day. RNA was extracted from two independent pools for each genotype.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Keiji Nakajima

PROVIDER: E-GEOD-30309 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The Arabidopsis RWP-RK protein RKD4 triggers gene expression and pattern formation in early embryogenesis.

Waki Takamitsu T Hiki Takeshi T Watanabe Ryouhei R Hashimoto Takashi T Nakajima Keiji K

Current biology : CB 20110728 15

Morphogenesis of seed plants commences with highly stereotypical cell division sequences in early embryogenesis [1, 2]. Although a small number of transcription factors and a mitogen-activated protein (MAP) kinase cascade have been implicated in this process [3-8], pattern formation in early embryogenesis remains poorly understood. We show here that the Arabidopsis RKD4, a member of the RWP-RK motif-containing putative transcription factors [9], is required for this process. Loss-of-function rkd ...[more]

PMID: 21802301

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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

Gene expression changes in response to the overexpression of the Arabidopsis RKD4 gene

Publications

The Arabidopsis RWP-RK protein RKD4 triggers gene expression and pattern formation in early embryogenesis.

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