HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: M500007-MCP200v1 4/10/1558    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Feilner, T. Articles by Kersten, B. Search for Related Content PubMed PubMed Citation Articles by Feilner, T. Articles by Kersten, B. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 4:1558-1568, 2005.
© 2005 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Research

High Throughput Identification of Potential Arabidopsis Mitogen-activated Protein Kinases Substrates^*,S

Tanja Feilner^,, Claus Hultschig, Justin Lee^¶, Svenja Meyer^||, Richard G. H. Immink^**, Andrea Koenig, Alexandra Possling^,, Harald Seitz, Allan Beveridge^,, Dierk Scheel^¶, Dolores J. Cahill^,, Hans Lehrach, Jürgen Kreutzberger and Birgit Kersten^,

From the Department of Vertebrate Genomics, Max Planck Institute for Molecular Genetics (MPI-MG), Ihnestr. 73, 14195 Berlin, Germany, ^¶ Department of Stress and Developmental Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120 Halle, Germany, ^|| RZPD German Resource Centre for Genome Research GmbH, Heubnerweg 6, 14059 Berlin, Germany, gabi.rzpd.de/, and ^** Business Unit Bioscience, Plant Research International, P O. Box 16, 6700 AA Wageningen, The Netherlands

Mitogen-activated protein kinase (MAPK) cascades are universal and highly conserved signal transduction modules in eucaryotes, including plants. These protein phosphorylation cascades link extracellular stimuli to a wide range of cellular responses. However, the underlying mechanisms are so far unknown as information about phosphorylation substrates of plant MAPKs is lacking. In this study we addressed the challenging task of identifying potential substrates for Arabidopsis thaliana mitogen-activated protein kinases MPK3 and MPK6, which are activated by many environmental stress factors. For this purpose, we developed a novel protein microarray-based proteomic method allowing high throughput study of protein phosphorylation. We generated protein microarrays including 1,690 Arabidopsis proteins, which were obtained from the expression of an almost nonredundant uniclone set derived from an inflorescence meristem cDNA expression library. Microarrays were incubated with MAPKs in the presence of radioactive ATP. Using a threshold-based quantification method to evaluate the microarray results, we were able to identify 48 potential substrates of MPK3 and 39 of MPK6. 26 of them are common for both kinases. One of the identified MPK6 substrates, 1-aminocyclopropane-1-carboxylic acid synthase-6, was just recently shown as the first plant MAPK substrate in vivo, demonstrating the potential of our method to identify substrates with physiological relevance. Furthermore we revealed transcription factors, transcription regulators, splicing factors, receptors, histones, and others as candidate substrates indicating that regulation in response to MAPK signaling is very complex and not restricted to the transcriptional level. Nearly all of the 48 potential MPK3 substrates were confirmed by other in vitro methods. As a whole, our approach makes it possible to shortlist candidate substrates of mitogen-activated protein kinases as well as those of other protein kinases for further analysis. Follow-up in vivo experiments are essential to evaluate their physiological relevance.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				G. J.M. Beckers, M. Jaskiewicz, Y. Liu, W. R. Underwood, S. Y. He, S. Zhang, and U. Conrath Mitogen-Activated Protein Kinases 3 and 6 Are Required for Full Priming of Stress Responses in Arabidopsis thaliana PLANT CELL, March 1, 2009; 21(3): 944 - 953. [Abstract] [Full Text] [PDF]

				S. C. Popescu, G. V. Popescu, S. Bachan, Z. Zhang, M. Gerstein, M. Snyder, and S. P. Dinesh-Kumar MAPK target networks in Arabidopsis thaliana revealed using functional protein microarrays Genes & Dev., January 1, 2009; 23(1): 80 - 92. [Abstract] [Full Text] [PDF]

				G. Merkouropoulos, E. Andreasson, D. Hess, T. Boller, and S. C. Peck An Arabidopsis Protein Phosphorylated in Response to Microbial Elicitation, AtPHOS32, Is a Substrate of MAP Kinases 3 and 6 J. Biol. Chem., April 18, 2008; 283(16): 10493 - 10499. [Abstract] [Full Text] [PDF]

				I. J.E. Stulemeijer, J. W. Stratmann, and M. H.A.J. Joosten Tomato Mitogen-Activated Protein Kinases LeMPK1, LeMPK2, and LeMPK3 Are Activated during the Cf-4/Avr4-Induced Hypersensitive Response and Have Distinct Phosphorylation Specificities Plant Physiology, July 1, 2007; 144(3): 1481 - 1494. [Abstract] [Full Text] [PDF]

				J. Wu, C. Hettenhausen, S. Meldau, and I. T. Baldwin Herbivory Rapidly Activates MAPK Signaling in Attacked and Unattacked Leaf Regions but Not between Leaves of Nicotiana attenuata PLANT CELL, March 1, 2007; 19(3): 1096 - 1122. [Abstract] [Full Text] [PDF]

				S. de la Fuente van Bentem, D. Anrather, E. Roitinger, A. Djamei, T. Hufnagl, A. Barta, E. Csaszar, I. Dohnal, D. Lecourieux, and H. Hirt Phosphoproteomics reveals extensive in vivo phosphorylation of Arabidopsis proteins involved in RNA metabolism Nucleic Acids Res., July 17, 2006; 34(11): 3267 - 3278. [Abstract] [Full Text] [PDF]

				A. Pitzschke and H. Hirt Mitogen-Activated Protein Kinases and Reactive Oxygen Species Signaling in Plants Plant Physiology, June 1, 2006; 141(2): 351 - 356. [Full Text] [PDF]

				G. Grelle, S. Kostka, A. Otto, B. Kersten, K. F. Genser, E.-C. Muller, S. Walter, A. Boddrich, U. Stelzl, C. Hanig, et al. Identification of VCP/p97, Carboxyl Terminus of Hsp70-interacting Protein (CHIP), and Amphiphysin II Interaction Partners Using Membrane-based Human Proteome Arrays Mol. Cell. Proteomics, February 1, 2006; 5(2): 234 - 244. [Abstract] [Full Text] [PDF]