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Subcellular Localization and Oligomerization of the Arabidopsis thaliana Somatic Embryogenesis Receptor Kinase 1 Protein

Citation

Shad, K and Gadella, TW and van Erp, H and Hecht, VFG and de Vries, SC, Subcellular Localization and Oligomerization of the Arabidopsis thaliana Somatic Embryogenesis Receptor Kinase 1 Protein, Journal of Molecular Biology, 309, (3) pp. 641-655. ISSN 0022-2836 (2001) [Refereed Article]

DOI: doi:10.1006/jmbi.2001.4706

Abstract

The Arabidopsis thaliana somatic embryogenesis receptor kinase 1 (AtSERK1) gene is expressed in developing ovules and early embryos. AtSERK1 is also transiently expressed during somatic embryogenesis. The predicted AtSERK1 protein contains an extracellular domain with a leucine zipper motif followed by five leucine-rich repeats, a proline-rich region, a single transmembrane region and an intracellular kinase domain. The AtSERK1 cDNA was fused to two different variants of green fluorescent protein (GFP), a yellow-emittin g GFP (YFP) and a cyan-emitting GFP (CFP), and transiently expressed in both plant protoplasts and insect cells. Using confocal laser scanning microscopy it was determined that the AtSERK1-YFP fusion protein is targeted to plasma membranes in both plant and animal cells. The extracellular leucine-rich repeats, and in particular the N-linked oligosaccharides that are present on them appear to be essential for correct localization of the AtSERK1-YFP protein. The potential for dimerization of the AtSERK1 protein was investigated by measuring the YFP/CFP fluorescence emission ratio using fluorescence spectral imaging microscopy. This ratio will increase due to fluorescence resonance energy transfer if the AtSERK1-CFP and AtSERK1-YFP fusion proteins interact. In 15% of the cells the YFP/CFP emission ratio for plasma membrane localized AtSERK1 proteins was enhanced. Yeast-protein interaction experiments confirmed the possibility for AtSERK1 homodimerization. Elimination of the extracellular leucine zipper domain reduced the YFP/CFP emission ratio to control levels indicating that without the leucine zipper domain AtSERK1 is monomeric. © 2001 Academic Press.

Item Details

Item Type:Refereed Article
Research Division:Biological Sciences
Research Group:Genetics
Research Field:Gene Expression (incl. Microarray and other genome-wide approaches)
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Environmental Sciences
Author:Hecht, VFG (Dr Valerie Hecht)
ID Code:48234
Year Published:2001
Web of Science® Times Cited:50
Deposited By:Plant Science
Deposited On:2007-10-03
Last Modified:2011-11-08
Downloads:0

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