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First published online 5 October 2005
doi: 10.1242/dev.02055
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1 Department of Biology, Yonsei University, Sinchon 134, Seoul 120-749,
Korea
2 Division of Molecular and Life Sciences, Pohang University of Science and
Technology, Pohang 790-784, Korea
3 Department of Molecular, Cellular, and Developmental Biology, University of
Michigan, Ann Arbor, MI 48109, USA
* Author for correspondence (e-mail: mmlee{at}yonsei.ac.kr)
Accepted 23 August 2005
The Arabidopsis root epidermis is composed of two types of cells, hair cells and non-hair cells, and their fate is determined in a position-dependent manner. WEREWOLF (WER), a R2R3 MYB protein, has been shown genetically to function as a master regulator to control both of the epidermal cell fates. To directly test the proposed role of WER in this system, we examined its subcellular localization and defined its transcriptional activation properties. We show that a WER-GFP fusion protein is functional and accumulates in the nucleus of the N-position cells in the Arabidopsis root epidermis, as expected for a transcriptional regulator. We also find that a modified WER protein with a strong activation domain (WER-VP16) promotes the formation of both epidermal cell types, supporting the view that WER specifies both cell fates. In addition, we used the glucocorticoid receptor (GR) inducible system to show that CPC transcription is regulated directly by WER. Using EMSA, we found two WER-binding sites (WBSs; WBSI and WBSII) in the CPC promoter. WER-WBSI binding was confirmed in vivo using the yeast one-hybrid assay. Binding between the WER protein and both WBSs (WBSI and WBSII), and the importance of the two WBSs in CPC promoter activity were confirmed in Arabidopsis. These results provide experimental support for the proposed role of WER as an activator of gene transcription during the specification of both epidermal cell fates.
Key words: WEREWOLF, CAPRICE, EMSA, Direct target, Root epidermis, Cell fate specification, Arabidopsis
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