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Characterization of histidine-aspartate kinase HK1 and identification of histidine phosphotransfer proteins as potential partners in a Populus multistep phosphorelay

Authors

  • François Héricourt,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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    • These authors equally contributed to this work.

  • Françoise Chefdor,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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    • These authors equally contributed to this work.

  • Lucie Bertheau,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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  • Mirai Tanigawa,

    1. Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan
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  • Tatsuya Maeda,

    1. Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan
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  • Grégory Guirimand,

    1. Université François Rabelais de Tours, EA 2106, Biomolécules et Biotechnologies Végétales (BBV), Tours, France
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  • Vincent Courdavault,

    1. Université François Rabelais de Tours, EA 2106, Biomolécules et Biotechnologies Végétales (BBV), Tours, France
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  • Mélanie Larcher,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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  • Christiane Depierreux,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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  • Hélène Bénédetti,

    1. Centre de Biophysique Moléculaire, Orléans Cedex 2, France
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  • Domenico Morabito,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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  • Franck Brignolas,

    1. Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
    2. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
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  • Sabine Carpin

    Corresponding author
    1. INRA, USC1328, Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), Orléans Cedex 2, France
    • Université d'Orléans, UPRES EA 1207, Laboratoire de Biologie des Ligneux et des Grandes Cultures (LBLGC), Orléans Cedex 2, France
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Correspondence

Corresponding author,

e-mail: sabine.carpin@univ-orleans.fr

Abstract

In poplar, we identified proteins homologous to yeast proteins involved in osmosensing multistep phosphorelay Sln1p-Ypd1p-Ssk1p. This finding led us to speculate that Populus cells could sense osmotic stress by a similar mechanism. This study focuses on first and second protagonists of this possible pathway: a histidine-aspartate kinase (HK1), putative osmosensor and histidine phosphotransfer proteins (HPt1 to 10), potential partners of this HK. Characterization of HK1 showed its ability to homodimerize in two-hybrid tests and to act as an osmosensor with a kinase activity in yeast, by functional complementation of sln1Δ sho1Δ strain. Moreover, in plant cells, plasma membrane localization of HK1 is shown. Further analysis on HPts allowed us to isolate seven new cDNAs, leading to a total of 10 different HPts identified in poplar. Interaction tests showed that almost all HPts can interact with HK1, but two of them exhibit stronger interactions, suggesting a preferential partnership in poplar. The importance of the phosphorylation status in these interactions has been investigated with two-hybrid tests carried out with mutated HK1 forms. Finally, in planta co-expression analysis of genes encoding these potential partners revealed that only three HPts are co-expressed with HK1 in different poplar organs. This result reinforces the hypothesis of a partnership between HK1 and these three preferential HPts in planta. Taken together, these results shed some light on proteins partnerships that could be involved in the osmosensing pathway in Populus.

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