OPEN ACCESS ARTICLE

This Article Free via Open Access: OA OA Full Text Full Text (PDF) OA All Versions of this Article: 143/2/670    most recent pp.106.090159v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Related articles in Plant Physiol. Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (4) Citing Articles via Google Scholar Google Scholar Articles by Schoonheim, P. J. Articles by de Boer, A. H. Search for Related Content PubMed PubMed Citation Articles by Schoonheim, P. J. Articles by de Boer, A. H. Agricola Articles by Schoonheim, P. J. Articles by de Boer, A. H.

CELL BIOLOGY AND SIGNAL TRANSDUCTION

A Comprehensive Analysis of the 14-3-3 Interactome in Barley Leaves Using a Complementary Proteomics and Two-Hybrid Approach^1,[C],[OA]

Department of Structural Biology, Faculty of Earth and Life Sciences, Vrije Universiteit, 1081 HV Amsterdam, The Netherlands (P.J.S., H.V., D.d.C.P., A.H.d.B.); and Department of Plant Biology, Cornell University, Ithaca, New York 14853 (G.F., K.J.v.W.)

This study describes the identification of over 150 target proteins of the five 14-3-3 isoforms in 7-d-old barley (Hordeum vulgare) cv Himalaya seedlings using yeast two-hybrid screens complemented with 14-3-3 protein affinity purification and tandem mass spectrometry. Independent experiments for a subset of genes confirmed the yeast two-hybrid interactions, demonstrating a low false positive identification rate. These combined approaches resulted in the identification of more than 150 putative targets; 15% were previously reported to be 14-3-3 interactors, including, for example, Serpin, RF2A, WPK4 kinase, P-type proton-translocating adenosine triphosphatase, EF1A, glutamine synthetase, and invertases. The affinity purification resulted in 30 interactors, of which 44% function in metabolism, while the yeast two-hybrid screens identified 132 different proteins, with 35% of the proteins involved in signal transduction. A number of proteins have a well-described function in hormonal signaling, such as the auxin transport protein PIN1 and NPH3 and components of the brassinosteroid pathway, such as the receptor kinase BAK1 (OsPERK1) and BRI1-kinase domain-interacting protein 129. However, 14-3-3 interactions with these signal mediators have not been confirmed in the affinity purification. Confirmations of the 14-3-3 interaction with the three ABF-like transcription factors are shown using far western analysis. Also, a REPRESSION OF SHOOT GROWTH ortholog named RF2A was identified; these transcription factors play important roles in the abscisic acid and gibberellin pathways, respectively. We speculate that 14-3-3 proteins have a role in cross talk between these hormonal pathways. The specificity and complementary nature of both the affinity purification and the yeast two-hybrid approaches is discussed.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Albertus H. de Boer (bert.de.boer{at}falw.vu.nl).

^[C] Some figures in this article are displayed in color online but in black and white in the print edition.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.106.090159

^* Corresponding author; e-mail bert.de.boer{at}falw.vu.nl; fax 31205987136.

Received September 21, 2006; accepted December 6, 2006; published December 15, 2006.

Related articles in Plant Physiol.:

On the Inside

Peter V. Minorsky
Plant Physiol. 2007 143: 553-554. [Full Text]  

This article has been cited by other articles:

				A. Wasilewska, F. Vlad, C. Sirichandra, Y. Redko, F. Jammes, C. Valon, N. F. d. Frey, and J. Leung An Update on Abscisic Acid Signaling in Plants and More ... Mol Plant, March 1, 2008; 1(2): 198 - 217. [Abstract] [Full Text] [PDF]

				Z. Deng, X. Zhang, W. Tang, J. A. Oses-Prieto, N. Suzuki, J. M. Gendron, H. Chen, S. Guan, R. J. Chalkley, T. K. Peterman, et al. A Proteomics Study of Brassinosteroid Response in Arabidopsis Mol. Cell. Proteomics, December 1, 2007; 6(12): 2058 - 2071. [Abstract] [Full Text] [PDF]

				H. Ryu, K. Kim, H. Cho, J. Park, S. Choe, and I. Hwang Nucleocytoplasmic Shuttling of BZR1 Mediated by Phosphorylation Is Essential in Arabidopsis Brassinosteroid Signaling PLANT CELL, September 1, 2007; 19(9): 2749 - 2762. [Abstract] [Full Text] [PDF]

				M.-Y. Bai, L.-Y. Zhang, S. S. Gampala, S.-W. Zhu, W.-Y. Song, K. Chong, and Z.-Y. Wang Functions of OsBZR1 and 14-3-3 proteins in brassinosteroid signaling in rice PNAS, August 21, 2007; 104(34): 13839 - 13844. [Abstract] [Full Text] [PDF]