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Gene Networks and Chromatin and Transcriptional Regulation of the Phaseolin Promoter in Arabidopsis
Sabarinath Sundaram, Sunee Kertbundit, Eugene V. Shakirov, Lakshminarayan M. Iyer, Miloslav Juříček and Timothy C. Hall
The Plant Cell
Vol. 25, No. 7 (JULY 2013), pp. 2601-2617
Published by: American Society of Plant Biologists (ASPB)
Stable URL: http://www.jstor.org/stable/23598296
Page Count: 17
You can always find the topics here!Topics: Genes, Chromatin, Plants, Gene expression, Gene expression regulation, Plant cells, Datasets, RNA, Seed development, DNA
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The complete lack of seed storage protein expression in vegetative tissues and robust expression during embryogenesis makes seed development an ideal system to study tissue-specific expression of genes. The promoter for the Phaseolin (phas) gene, which encodes the major seed storage protein in bean (Phaseolus vulgaris), is activated in two sequential steps: Phaseolus vulgaris ABI3-like factor (Pv-ALF)—dependent potentiation and abscisic acid—mediated activation. In this study, a heterologous in vivo Pv-ALF/phas-GUS (for β-glucuronidase) expression system in transgenic Arabidopsis thaliana leaves was used in conjunction with the powerful RNA-Seq approach to capture transcriptional landscapes of phas promoter expression. Remarkably, expression of over 1300 genes from 11 functional categories coincided with changes in the transcriptional status of the phas promoter. Gene network analysis of induced genes and artificial microRNA—mediated loss-of-function genetic assays identified transcriptional regulators RINGLET 2 (RLT2) and AINTEGUMENTA-LIKE 5 (AIL5) as being essential for phas transcription. Pv-ALF binding to the RLT2 and AIL5 promoter regions was confirmed by electrophoretic mobility shift assay. RLT2 and AIL5 knockdown lines displayed reduced expression of several endogenous seed genes, suggesting that these factors are involved in activation of endogenous Arabidopsis seed storage gene expression. Overall, the identification of these key factors involved in phas activation provides important insight into the two-step transcriptional regulation of seed-specific gene expression.
The Plant Cell © 2013 American Society of Plant Biologists (ASPB)