ABA Signaling in Guard Cells Entails a Dynamic Protein–Protein Interaction Relay from the PYL-RCAR Family Receptors to Ion Channels
ABSTRACT Plant hormone abscisic acid (ABA) serves as an integrator of environmental stresses such as drought to trigger stomatal closure by regulating specific ion channels in guard cells. We previously reported that SLAC1, an outward anion channel required for stomatal closure, was regulated via reversible protein phosphorylation events involving ABA signaling components, including protein phosphatase 2C members and a SnRK2-type kinase (OST1). In this study, we reconstituted the ABA signaling pathway as a protein-protein interaction relay from the PYL/RCAR-type receptors, to the PP2C-SnRK2 phosphatase-kinase pairs, to the ion channel SLAC1. The ABA receptors interacted with and inhibited PP2C phosphatase activity against the SnRK2-type kinase, releasing active SnRK2 kinase to phosphorylate, and activate the SLAC1 channel, leading to reduced guard cell turgor and stomatal closure. Both yeast two-hybrid and bimolecular fluorescence complementation assays were used to verify the interactions among the components in the pathway. These biochemical assays demonstrated activity modifications of phosphatases and kinases by their interaction partners. The SLAC1 channel activity was used as an endpoint readout for the strength of the signaling pathway, depending on the presence of different combinations of signaling components. Further study using transgenic plants overexpressing one of the ABA receptors demonstrated that changing the relative level of interacting partners would change ABA sensitivity.
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- "Ten leaves were detached from each plant and placed in petridishes , and the loss of fresh weight was determined at the indicated times. Stomatal aperture bioassay was carried out as described before with the following modifications (Lee et al., 2013). In briefly, four rosette leaves of 4-week-old plants were detached and floated in stomatal opening solution (SOS: 50 mM KCl and 10 mM MES-KOH, pH 6.15, 10 µM CaCl 2 ) in the light. "
ABSTRACT: Plants are frequently exposed to numerous environmental stresses such as dehydration and high salinity, and have developed elaborate mechanisms to counteract the deleterious effects of stress. The phytohormone abscisic acid (ABA) plays a critical role as an integrator of plant responses to water-limited condition to activate ABA signal transduction pathway. Although perception of ABA has been suggested to be important, the function of each ABA receptor remains elusive in dehydration condition. Here, we show that ABA receptor, pyrabactin resistance-like protein 8 (PYL8), functions in dehydration conditions. Transgenic plants overexpressing PYL8 exhibited hypersensitive phenotype to ABA in seed germination, seedling growth and establishment. We found that hypersensitivity to ABA of transgenic plants results in high degrees of stomatal closure in response to ABA leading to low transpiration rates and ultimately more vulnerable to drought than the wild-type plants. In addition, high expression of ABA maker genes also contributes to altered drought tolerance phenotype. Overall, this work emphasizes the importance of ABA signaling by ABA receptor in stomata during defense response to drought stress.The plant pathology journal 12/2013; 29(4). DOI:10.5423/PPJ.NT.07.2013.0071 · 0.76 Impact Factor
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ABSTRACT: Abscisic acid (ABA) is an essential molecule in plant abiotic stress responses. It binds to soluble pyrabactin resistance1/PYR1-like/regulatory component of ABA receptor receptors and stabilizes them in a conformation that inhibits clade A type II C protein phosphatases; this leads to downstream SnRK2 kinase activation and numerous cellular outputs. We previously described the synthetic naphthalene sulfonamide ABA agonist pyrabactin, which activates seed ABA responses but fails to trigger substantial responses in vegetative tissues in Arabidopsis thaliana. Here we describe quinabactin, a sulfonamide ABA agonist that preferentially activates dimeric ABA receptors and possesses ABA-like potency in vivo. In Arabidopsis, the transcriptional responses induced by quinabactin are highly correlated with those induced by ABA treatments. Quinabactin treatments elicit guard cell closure, suppress water loss, and promote drought tolerance in adult Arabidopsis and soybean plants. The effects of quinabactin are sufficiently similar to those of ABA that it is able to rescue multiple phenotypes observed in the ABA-deficient mutant aba2. Genetic analyses show that quinabactin's effects in vegetative tissues are primarily mediated by dimeric ABA receptors. A PYL2-quinabactin-HAB1 X-ray crystal structure solved at 1.98-Å resolution shows that quinabactin forms a hydrogen bond with the receptor/PP2C "lock" hydrogen bond network, a structural feature absent in pyrabactin-receptor/PP2C complexes. Our results demonstrate that ABA receptors can be chemically controlled to enable plant protection against water stress and define the dimeric receptors as key targets for chemical modulation of vegetative ABA responses.Proceedings of the National Academy of Sciences 07/2013; 110(29). DOI:10.1073/pnas.1305919110 · 9.81 Impact Factor
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ABSTRACT: Plants are frequently exposed to various environmental stresses including drought in the natural environment and have evolved physiological, biochemical, and molecular mechanisms to counteract the deleterious effects of stress. Of them, modulation of abscisic acid (ABA) signal transduction allows plants to overcome stress. Recently, Kim and Hwang (Plant J 72:843-855, 2012) identified CaMLO2 that is transcriptionally induced by both biotic and abiotic stress. Based on this, we tested the possibility that CaMLO2 is involved in abiotic stress, although m ildew resistance l ocus O (MLO) proteins have been known as negative regulators in plant defense responses against powdery mildew. The CaMLO2 gene was strongly induced in pepper leaves exposed to ABA and drought. Virus-induced gene silencing of CaMLO2 in pepper plants showed low levels of transpiration and lipid peroxidation in dehydrated leaves. Overexpression of the CaMLO2 gene in Arabidopsis conferred reduced sensitivity to ABA in germination and seedling growth and establishment. High transpiration rates and low degrees of stomatal closure in response to ABA also led transgenic plants to be more vulnerable to drought than the wild-type, which was accompanied by altered expression of stress-related genes. Taken together, these data suggest that CaMLO2 acts as a negative regulator of ABA signaling that suppresses water loss from leaves under drought conditions.Plant Molecular Biology 11/2013; 85(1-2). DOI:10.1007/s11103-013-0155-8 · 4.07 Impact Factor