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Free Content A fast brassinolide-regulated response pathway in the plasma membrane of Arabidopsis thaliana

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Summary

To understand molecular processes in living plant cells, quantitative spectro-microscopic technologies are required. By combining fluorescence lifetime spectroscopy with confocal microscopy, we studied the subcellular properties and function of a GFP-tagged variant of the plasma membrane-bound brassinosteroid receptor BRI1 (BRI1–GFP) in living cells of Arabidopsis seedlings. Shortly after adding brassinolide, we observed BRI1-dependent cell-wall expansion, preceding cell elongation. In parallel, the fluorescence lifetime of BRI1–GFP decreased, indicating an alteration in the receptor’s physico-chemical environment. The parameter modulating the fluorescence lifetime of BRI1–GFP was found to be BL-induced hyperpolarization of the plasma membrane. Furthermore, for induction of hyperpolarization and cell-wall expansion, activation of the plasma membrane P-ATPase was necessary. This activation required BRI1 kinase activity, and was mediated by BL-modulated interaction of BRI1 with the P-ATPase. Our results were used to develop a model suggesting that there is a fast BL-regulated signal response pathway within the plasma membrane that links BRI1 with P-ATPase for the regulation of cell-wall expansion.
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Keywords: P-ATPase; brassinolide; brassinosteroid-insensitive 1 (BRI1); cell wall; fluorescence lifetime; membrane potential

Document Type: Research Article

Affiliations: 1: Laboratory of Plant Physiology and Biophysics, Faculty of Biomedical and Life Sciences – Bower Building, University of Glasgow, Glasgow G12 8QQ, UK 2: Center for Plant Molecular Biology, Department of Plant Physiology, University of Tübingen, Auf der Morgenstelle 1, 72076 Tübingen, Germany

Publication date: May 1, 2011

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