Co-immunoprecipitation-based identification of putative BAX INHIBITOR-1-interacting proteins involved in cell death regulation and plant-powdery mildew interactions

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2013
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Weis, Corina
Pfeilmeier, Sebastian
Glawischnig, Erich
Pachl, Fiona
Hahne, Hannes
Kuster, Bernhard
Eichmann, Ruth
Hückelhoven, Ralph
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Molecular Plant Pathology. 2013, 14(8), pp. 791-802. eISSN 1464-6722. Available under: doi: 10.1111/mpp.12050
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The endoplasmic reticulum (ER)-resident BAX INHIBITOR-1 (BI-1) protein is one of a few cell death suppressors known to be conserved in animals and plants. The function of BI-1 proteins in response to various biotic and abiotic stress factors is well established. However, little is known about the underlying mechanisms. We conducted co-immunoprecipitation (co-IP) experiments to identify Arabidopsis thaliana BI-1-interacting proteins to obtain a potentially better understanding of how BI-1 functions during plant-pathogen interactions and as a suppressor of cell death. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 95 proteins co-immunoprecipitated with green fluorescing protein (GFP)-tagged BI-1. Five selected candidate proteins, a RIBOPHORIN II (RPN2) family protein, VACUOLAR ATP SYNTHASE SUBUNIT A (VHA-A), cytochrome P450 83A1 (CYP83A1), H+ -ATPASE 1 (AHA1) and PROHIBITIN 2 (PHB2), were further investigated with regard to their role in BI-1-associated processes. To this end, we analysed a set of Arabidopsis mutants in the interaction with the adapted powdery mildew fungus Erysiphe cruciferarum and on cell death-inducing treatments. Two independent rpn2 knock-down mutants tended to better support powdery mildew, and a phb2 mutant showed altered responses to cell death-inducing Alternaria alternata f.sp. lycopersici (AAL) toxin treatment. Two independent cyp83a1 mutants showed a strong powdery mildew resistance phenotype and enhanced sensitivity to AAL toxin. Moreover, co-localization studies and fluorescence resonance energy transfer (FRET) experiments suggested a direct interaction of BI-1 with CYP83A1 at the ER.

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ISO 690WEIS, Corina, Sebastian PFEILMEIER, Erich GLAWISCHNIG, Erika ISONO, Fiona PACHL, Hannes HAHNE, Bernhard KUSTER, Ruth EICHMANN, Ralph HÜCKELHOVEN, 2013. Co-immunoprecipitation-based identification of putative BAX INHIBITOR-1-interacting proteins involved in cell death regulation and plant-powdery mildew interactions. In: Molecular Plant Pathology. 2013, 14(8), pp. 791-802. eISSN 1464-6722. Available under: doi: 10.1111/mpp.12050
BibTex
@article{Weis2013-10Coimm-38395,
  year={2013},
  doi={10.1111/mpp.12050},
  title={Co-immunoprecipitation-based identification of putative BAX INHIBITOR-1-interacting proteins involved in cell death regulation and plant-powdery mildew interactions},
  number={8},
  volume={14},
  journal={Molecular Plant Pathology},
  pages={791--802},
  author={Weis, Corina and Pfeilmeier, Sebastian and Glawischnig, Erich and Isono, Erika and Pachl, Fiona and Hahne, Hannes and Kuster, Bernhard and Eichmann, Ruth and Hückelhoven, Ralph}
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