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Bimolecular Fluorescent Complementation (BiFC) by MAP Kinases and MAPK Phosphatases

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Plant MAP Kinases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1171))

Abstract

The adaptation of plants to the environment is a key property for survival. Adaptation responses to environmental cues are generated in cells by signaling initiated from cell receptors. Signal transduction is based on protein phosphorylation that is employed in mitogen-activated protein kinase (MAPK) cascades to integrate signals from receptors to cellular responses. MAPK activity is determined by phosphorylation of amino acid residues within the kinase activation loop and their dephosphorylation by phosphatases is essential to control signal duration and intensity.

Monitoring protein–protein interactions (PPIs) of MAPKs with MAPK phosphatases in vivo provides valuable information about specificity and intracellular localization of the protein complex. Here, we report studying PPIs between Arabidopsis MAPKs and PP2C-type MAPK phosphatases using bimolecular fluorescent complementation (BiFC) in suspension cell protoplasts. The interactions of the MAPKs MPK3, MKP4 and MPK6 with the phosphatases AP2C1 and AP2C3 have been tested.

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Acknowledgements

We thank Dierk Scheel and Thomas Kroj for sharing the protoplast isolation method for parsley and Andrej Belokurov for cultivation of suspension culture cells. This work has received funding from the Lithuanian-Swiss cooperation program to reduce economic and social disparities within the enlarged European Union under project agreement No CH-3-ŠMM-01/10 and from the Austrian Science Fund (FWF) I255, L687 and W1220-B09.

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Authors and Affiliations

  1. Max F. Perutz Laboratories, University and Medical University of Vienna, Dr. Bohrgasse 9, 1030, Vienna, Austria

    Alois Schweighofer, Volodymyr Shubchynskyy & Irute Meskiene

  2. Institute of Biotechnology (IBT), University of Vilnius, V. Graičiūno 8, 02241, Vilnius, Lithuania

    Alois Schweighofer, Vaiva Kazanaviciute & Irute Meskiene

  3. Gregor Mendel Institute of Molecular Plant Biology (GMI), Dr. Bohrgasse 3, 1030, Vienna, Austria

    Armin Djamei

Authors
  1. Alois Schweighofer
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  2. Volodymyr Shubchynskyy
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  3. Vaiva Kazanaviciute
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  4. Armin Djamei
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  5. Irute Meskiene
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Corresponding author

Correspondence to Alois Schweighofer .

Editor information

Editors and Affiliations

  1. Palacky University in Olomouc, Olomouc, Czech Republic

    George Komis

  2. Faculty of Science, Palacky University in Olomouc, Olomouc, Czech Republic

    Jozef Šamaj

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Schweighofer, A., Shubchynskyy, V., Kazanaviciute, V., Djamei, A., Meskiene, I. (2014). Bimolecular Fluorescent Complementation (BiFC) by MAP Kinases and MAPK Phosphatases. In: Komis, G., Šamaj, J. (eds) Plant MAP Kinases. Methods in Molecular Biology, vol 1171. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0922-3_12

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  • DOI: https://doi.org/10.1007/978-1-4939-0922-3_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0921-6

  • Online ISBN: 978-1-4939-0922-3

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