The Ca2+ Sensor SCaBP3/CBL7 Modulates Plasma Membrane H+-ATPase Activity and Promotes Alkali Tolerance in Arabidopsis.

Saline-alkali soil is a major environmental constraint impairing plant growth and crop productivity. In this study, we identified a Ca2+ sensor/kinase/plasma membrane (PM) H+-ATPase module as a central component conferring alkali tolerance in Arabidopsis (Arabidopsis thaliana). We report that the SCaBP3 (SOS3-LIKE CALCIUM BINDING PROTEIN3)/CBL7 (CALCINEURIN B-LIKE7) loss-of-function plants exhibit enhanced stress tolerance associated with increased PM H+-ATPase activity and provide fundamental mechanistic insights into the regulation of PM H+-ATPase activity. Consistent with the genetic evidence, interaction analyses, in vivo reconstitution experiments, and determination of H+-ATPase activity indicate that interaction of the Ca2+ sensor SCaBP3 with the C-terminal Region I domain of the PM H+-ATPase AHA2 (Arabidopsis thaliana PLASMA MEMBRANE PROTON ATPASE2) facilitates the intramolecular interaction of the AHA2 C terminus with the Central loop region of the PM H+-ATPase to promote autoinhibition of H+-ATPase activity. Concurrently, direct interaction of SCaPB3 with the kinase PKS5 (PROTEIN KINASE SOS2-LIKE5) stabilizes the kinase-ATPase interaction and thereby fosters the inhibitory phosphorylation of AHA2 by PKS5. Consistently, yeast reconstitution experiments and genetic analysis indicate that SCaBP3 provides a bifurcated pathway for coordinating intramolecular and intermolecular inhibition of PM H+-ATPase. We propose that alkaline stress-triggered Ca2+ signals induce SCaBP3 dissociation from AHA2 to enhance PM H+-ATPase activity. This work illustrates a versatile signaling module that enables the stress-responsive adjustment of plasma membrane proton fluxes.

[1]  M. Sussman,et al.  In vivo cross-linking supports a head-to-tail mechanism for regulation of the plant plasma membrane P-type H+-ATPase , 2018, The Journal of Biological Chemistry.

[2]  Yanming Zhu,et al.  GsSLAH3, a Glycine soja slow type anion channel homolog, positively modulates plant bicarbonate stress tolerance. , 2018, Physiologia plantarum.

[3]  Yan Guo,et al.  Unraveling salt stress signaling in plants. , 2018, Journal of integrative plant biology.

[4]  Nan Wang,et al.  Calcium Signaling during Salt Stress and in the Regulation of Ion Homeostasis. , 2018, Journal of experimental botany.

[5]  J. Kudla,et al.  Advances and current challenges in calcium signaling. , 2018, The New phytologist.

[6]  Kyle W. Bender,et al.  Revisiting paradigms of Ca2+ signaling protein kinase regulation in plants. , 2018, The Biochemical journal.

[7]  Yan Guo,et al.  Elucidating the molecular mechanisms mediating plant salt-stress responses. , 2018, The New phytologist.

[8]  Lucélia Borgo.  Evaluation of buffers toxicity in tobacco cells: Homopiperazine-1,4-bis (2-ethanesulfonic acid) is a suitable buffer for plant cells studies at low pH. , 2017, Plant physiology and biochemistry : PPB.

[9]  Jian‐Kang Zhu.  Abiotic Stress Signaling and Responses in Plants , 2016, Cell.

[10]  M. Palmgren,et al.  Plasma Membrane H(+)-ATPase Regulation in the Center of Plant Physiology. , 2016, Molecular plant.

[11]  T. Cuin,et al.  Receptor kinase-mediated control of primary active proton pumping at the plasma membrane. , 2014, The Plant journal : for cell and molecular biology.

[12]  J. Kudla,et al.  The vacuolar calcium sensors CBL2 and CBL3 affect seed size and embryonic development in Arabidopsis thaliana. , 2014, The Plant journal : for cell and molecular biology.

[13]  L. An,et al.  The CBL–CIPK network mediates different signaling pathways in plants , 2013, Plant Cell Reports.

[14]  Wei-Hua Wu,et al.  Calcineurin B-like protein CBL10 directly interacts with AKT1 and modulates K+ homeostasis in Arabidopsis. , 2013, The Plant journal : for cell and molecular biology.

[15]  Xin He,et al.  The Ca2+ -dependent DNases are involved in secondary xylem development in Eucommia ulmoides. , 2012, Journal of integrative plant biology.

[16]  Ken-ichiro Hayashi,et al.  Auxin Activates the Plasma Membrane H+-ATPase by Phosphorylation during Hypocotyl Elongation in Arabidopsis1[W][OA] , 2012, Plant Physiology.

[17]  J. Kudla,et al.  Phosphorylation of Calcineurin B-like (CBL) Calcium Sensor Proteins by Their CBL-interacting Protein Kinases (CIPKs) Is Required for Full Activity of CBL-CIPK Complexes toward Their Target Proteins* , 2012, The Journal of Biological Chemistry.

[18]  M. Sussman,et al.  The Effect of a Genetically Reduced Plasma Membrane Protonmotive Force on Vegetative Growth of Arabidopsis1[C][W][OA] , 2012, Plant Physiology.

[19]  O. Jensen,et al.  Phosphosite Mapping of P-type Plasma Membrane H+-ATPase in Homologous and Heterologous Environments* , 2011, The Journal of Biological Chemistry.

[20]  J. Kudla,et al.  Calcium-dependent modulation and plasma membrane targeting of the AKT2 potassium channel by the CBL4/CIPK6 calcium sensor/protein kinase complex , 2011, Cell Research.

[21]  Wenhua Zhang,et al.  Rice phospholipase Dα is involved in salt tolerance by the mediation of H(+)-ATPase activity and transcription. , 2011, Journal of integrative plant biology.

[22]  Dieter Braun,et al.  Protein-binding assays in biological liquids using microscale thermophoresis. , 2010, Nature communications.

[23]  M. Palmgren,et al.  A Novel Mechanism of P-type ATPase Autoinhibition Involving Both Termini of the Protein , 2010, The Journal of Biological Chemistry.

[24]  Leonie Steinhorst,et al.  CBL-mediated targeting of CIPKs facilitates the decoding of calcium signals emanating from distinct cellular stores. , 2009, The Plant journal : for cell and molecular biology.

[25]  S. Chen,et al.  The Arabidopsis Chaperone J3 Regulates the Plasma Membrane H+-ATPase through Interaction with the PKS5 Kinase[C][W] , 2010, Plant Cell.

[26]  Poul Nissen,et al.  A structural overview of the plasma membrane Na+,K+-ATPase and H+-ATPase ion pumps , 2010, Nature Reviews Molecular Cell Biology.

[27]  W. Schmidt,et al.  Dissecting iron deficiency-induced proton extrusion in Arabidopsis roots. , 2009, The New phytologist.

[28]  M. Palmgren,et al.  RIN4 Functions with Plasma Membrane H+-ATPases to Regulate Stomatal Apertures during Pathogen Attack , 2009, PLoS biology.

[29]  R. Derua,et al.  Activation of Plant Plasma Membrane H+-ATPase by 14-3-3 Proteins Is Negatively Controlled by Two Phosphorylation Sites within the H+-ATPase C-terminal Region* , 2009, Journal of Biological Chemistry.

[30]  J. Kudla,et al.  Multicolor bimolecular fluorescence complementation reveals simultaneous formation of alternative CBL/CIPK complexes in planta. , 2008, The Plant journal : for cell and molecular biology.

[31]  P. Nissen,et al.  Crystal structure of the plasma membrane proton pump , 2008, Nature.

[32]  S. Merlot,et al.  Constitutive activation of a plasma membrane H+‐ATPase prevents abscisic acid‐mediated stomatal closure , 2007, The EMBO journal.

[33]  Rongmin Zhao,et al.  Expression of a Constitutively Activated Plasma Membrane H+-ATPase Alters Plant Development and Increases Salt Tolerance1[C][OA] , 2007, Plant Physiology.

[34]  Jian-Kang Zhu,et al.  Arabidopsis Protein Kinase PKS5 Inhibits the Plasma Membrane H+-ATPase by Preventing Interaction with 14-3-3 Protein , 2007, The Plant Cell Online.

[35]  C. Ottmann,et al.  Structure of a 14-3-3 coordinated hexamer of the plant plasma membrane H+ -ATPase by combining X-ray crystallography and electron cryomicroscopy. , 2007, Molecular cell.

[36]  Wei-Hua Wu,et al.  A Protein Kinase, Interacting with Two Calcineurin B-like Proteins, Regulates K+ Transporter AKT1 in Arabidopsis , 2006, Cell.

[37]  Sergio Marco,et al.  Activation of the plant plasma membrane H+-ATPase by phosphorylation and binding of 14-3-3 proteins converts a dimer into a hexamer. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[38]  Xiao-jing Wang,et al.  [Effects of methyl jasmonate treatment on the hydrolytic activity and phosphorylation level of plasma membrane H+-ATPase in mung bean (Vigna radiata L.) hypocotyls]. , 2004, Zhi wu sheng li yu fen zi sheng wu xue xue bao = Journal of plant physiology and molecular biology.

[39]  Klaus Harter,et al.  Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation. , 2004, The Plant journal : for cell and molecular biology.

[40]  A. Stensballe,et al.  Phosphoproteomics of the Arabidopsis Plasma Membrane and a New Phosphorylation Site Databasew⃞ , 2004, The Plant Cell Online.

[41]  M. Sussman,et al.  Phosphorylation of the plasma-membrane H+-ATPase of oat roots by a calcium-stimulated protein kinase , 1988, Planta.

[42]  E. Wagner,et al.  Plasma Membrane H+-ATPase Is Involved in Auxin-Mediated Cell Elongation during Wheat Embryo Development1 , 2003, Plant Physiology.

[43]  M. Bonza,et al.  A novel interaction partner for the C-terminus of Arabidopsis thaliana plasma membrane H+-ATPase (AHA1 isoform): site and mechanism of action on H+-ATPase activity differ from those of 14-3-3 proteins. , 2002, The Plant journal : for cell and molecular biology.

[44]  M. Palmgren,et al.  Post-translational Modification of Plant Plasma Membrane H+-ATPase as a Requirement for Functional Complementation of a Yeast Transport Mutant* , 2002, The Journal of Biological Chemistry.

[45]  J. Rudd,et al.  Unravelling response-specificity in Ca2+ signalling pathways in plant cells. , 2001, The New phytologist.

[46]  M. Palmgren.  PLANT PLASMA MEMBRANE H+-ATPases: Powerhouses for Nutrient Uptake. , 2001, Annual review of plant physiology and plant molecular biology.

[47]  R. Turgeon,et al.  Identification of phloem involved in assimilate loading in leaves by the activity of the galactinol synthase promoter. , 2000, Plant physiology.

[48]  J Vandekerckhove,et al.  A Plant Plasma Membrane H+-ATPase Expressed in Yeast Is Activated by Phosphorylation at Its Penultimate Residue and Binding of 14-3-3 Regulatory Proteins in the Absence of Fusicoccin* , 2000, The Journal of Biological Chemistry.

[49]  A. Stensballe,et al.  Binding of 14-3-3 Protein to the Plasma Membrane H+-ATPase AHA2 Involves the Three C-terminal Residues Tyr946-Thr-Val and Requires Phosphorylation of Thr947 * , 1999, The Journal of Biological Chemistry.

[50]  K. Axelsen,et al.  Molecular dissection of the C-terminal regulatory domain of the plant plasma membrane H+-ATPase AHA2: mapping of residues that when altered give rise to an activated enzyme. , 1999, Biochemistry.

[51]  M. Pessarakli.  Handbook of plant and crop stress , 1999 .

[52]  O. Maudoux,et al.  Single Point Mutations Distributed in 10 Soluble and Membrane Regions of the Nicotiana plumbaginifolia Plasma Membrane PMA2 H+-ATPase Activate the Enzyme and Modify the Structure of the C-terminal Region* , 1998, The Journal of Biological Chemistry.

[53]  A. Goffeau,et al.  Single point mutations in various domains of a plant plasma membrane H(+)‐ATPase expressed in Saccharomyces cerevisiae increase H(+)‐pumping and permit yeast growth at low pH. , 1996, The EMBO journal.

[54]  E. Blumwald,et al.  Regulation of Plant Defense Response to Fungal Pathogens: Two Types of Protein Kinases in the Reversible Phosphorylation of the Host Plasma Membrane H+-ATPase. , 1996, The Plant cell.

[55]  B. Regenberg,et al.  C-terminal deletion analysis of plant plasma membrane H(+)-ATPase: yeast as a model system for solute transport across the plant plasma membrane. , 1995, The Plant cell.

[56]  T. Kinoshita,et al.  Cytosolic Concentration of Ca2+ Regulates the Plasma Membrane H+-ATPase in Guard Cells of Fava Bean. , 1995, The Plant cell.

[57]  G. Bertoni,et al.  Alkalinity-bicarbonate-calcium effects on iron chlorosis in white lupine in soilless culture , 1995 .

[58]  A. Goffeau,et al.  A second transport ATPase gene in Saccharomyces cerevisiae. , 1988, The Journal of biological chemistry.

[59]  S. Noji,et al.  Effect of intracellular pH and potassium ions on a primary transport system for glutamate/aspartate in Streptococcus mutans. , 1988, European journal of biochemistry.

[60]  John A. Lee,et al.  A COMPARATIVE STUDY OF BICARBONATE INHIBITION OF ROOT GROWTH IN CALCICOLE AND CALCIFUGE GRASSES , 1969 .

[61]  The Binding Site for Regulatory 14-3-3 Protein in Plant Plasma Membrane H (cid:1) -ATPase INVOLVEMENT OF A REGION PROMOTING PHOSPHORYLATION-INDEPENDENT INTERACTION IN ADDITION TO THE PHOSPHORYLATION-DEPENDENT C-TERMINAL END* , 2022 .