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      A RAF-SnRK2 kinase cascade mediates early osmotic stress signaling in higher plants

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          Abstract

          Osmoregulation is important for plant growth, development and response to environmental changes. SNF1-related protein kinase 2s (SnRK2s) are quickly activated by osmotic stress and are central components in osmotic stress and abscisic acid (ABA) signaling pathways; however, the upstream components required for SnRK2 activation and early osmotic stress signaling are still unknown. Here, we report a critical role for B2, B3 and B4 subfamilies of Raf-like kinases (RAFs) in early osmotic stress as well as ABA signaling in Arabidopsis thaliana. B2, B3 and B4 RAFs are quickly activated by osmotic stress and are required for phosphorylation and activation of SnRK2s. Analyses of high-order mutants of RAFs reveal critical roles of the RAFs in osmotic stress tolerance and ABA responses as well as in growth and development. Our findings uncover a kinase cascade mediating osmoregulation in higher plants.

          Abstract

          Rapid activation of SnRK2 kinases is central to plant responses to osmotic stress and abscisic acid. Here the authors show that a group of Raf-like kinases are very quickly activated by osmotic stress, and then phosphorylate and activate SnRK2s.

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          Most cited references22

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          Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL family of START proteins.

          Type 2C protein phosphatases (PP2Cs) are vitally involved in abscisic acid (ABA) signaling. Here, we show that a synthetic growth inhibitor called pyrabactin functions as a selective ABA agonist. Pyrabactin acts through PYRABACTIN RESISTANCE 1 (PYR1), the founding member of a family of START proteins called PYR/PYLs, which are necessary for both pyrabactin and ABA signaling in vivo. We show that ABA binds to PYR1, which in turn binds to and inhibits PP2Cs. We conclude that PYR/PYLs are ABA receptors functioning at the apex of a negative regulatory pathway that controls ABA signaling by inhibiting PP2Cs. Our results illustrate the power of the chemical genetic approach for sidestepping genetic redundancy.
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            Regulators of PP2C phosphatase activity function as abscisic acid sensors.

            The plant hormone abscisic acid (ABA) acts as a developmental signal and as an integrator of environmental cues such as drought and cold. Key players in ABA signal transduction include the type 2C protein phosphatases (PP2Cs) ABI1 and ABI2, which act by negatively regulating ABA responses. In this study, we identify interactors of ABI1 and ABI2 which we have named regulatory components of ABA receptor (RCARs). In Arabidopsis, RCARs belong to a family with 14 members that share structural similarity with class 10 pathogen-related proteins. RCAR1 was shown to bind ABA, to mediate ABA-dependent inactivation of ABI1 or ABI2 in vitro, and to antagonize PP2C action in planta. Other RCARs also mediated ABA-dependent regulation of ABI1 and ABI2, consistent with a combinatorial assembly of receptor complexes.
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              Three SnRK2 protein kinases are the main positive regulators of abscisic acid signaling in response to water stress in Arabidopsis.

              Responses to water stress are thought to be mediated by transcriptional regulation of gene expression via reversible protein phosphorylation events. Previously, we reported that bZIP (basic-domain leucine zipper)-type AREB/ABF (ABA-responsive element-binding protein/factor) transcription factors are involved in ABA signaling under water stress conditions in Arabidopsis. The AREB1 protein is phosphorylated in vitro by ABA-activated SNF1-related protein kinase 2s (SnRK2s) such as SRK2D/SnRK2.2, SRK2E/SnRK2.6 and SRK2I/SnRK2.3 (SRK2D/E/I). Consistent with this, we now show that SRK2D/E/I and AREB1 co-localize and interact in nuclei in planta. Our results show that unlike srk2d, srk2e and srk2i single and double mutants, srk2d srk2e srk2i (srk2d/e/i) triple mutants exhibit greatly reduced tolerance to drought stress and highly enhanced insensitivity to ABA. Under water stress conditions, ABA- and water stress-dependent gene expression, including that of transcription factors, is globally and drastically impaired, and jasmonic acid (JA)-responsive and flowering genes are up-regulated in srk2d/e/i triple mutants, but not in other single and double mutants. The down-regulated genes in srk2d/e/i and areb/abf triple mutants largely overlap in ABA-dependent expression, supporting the view that SRK2D/E/I regulate AREB/ABFs in ABA signaling in response to water stress. Almost all dehydration-responsive LEA (late embryogenesis abundant) protein genes and group-A PP2C (protein phosphatase 2C) genes are strongly down-regulated in the srk2d/e/i triple mutants. Further, our data show that these group-A PP2Cs, such as HAI1 and ABI1, interact with SRK2D. Together, our results indicate that SRK2D/E/I function as main positive regulators, and suggest that ABA signaling is controlled by the dual modulation of SRK2D/E/I and group-A PP2Cs.
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                Author and article information

                Contributors
                jkzhu@sibs.ac.cn
                pcwang@sibs.ac.cn
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                30 January 2020
                30 January 2020
                2020
                : 11
                : 613
                Affiliations
                [1 ]ISNI 0000000119573309, GRID grid.9227.e, Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, , Chinese Academy of Sciences, ; Shanghai, 200032 China
                [2 ]ISNI 0000 0004 0530 8290, GRID grid.22935.3f, State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, , China Agricultural University, ; Beijing, 100193 China
                [3 ]ISNI 0000 0004 1937 2197, GRID grid.169077.e, Department of Horticulture and Landscape Architecture, , Purdue University, ; West Lafayette, IN 47907 USA
                [4 ]ISNI 0000 0000 9139 560X, GRID grid.256922.8, Key laboratory of Plant Stress Biology, School of Life Sciences, , Henan University, ; Kaifeng, 475004 China
                [5 ]ISNI 0000 0004 1937 2197, GRID grid.169077.e, Department of Biochemistry, , Purdue University, ; West Lafayette, IN 47907 USA
                [6 ]ISNI 0000 0004 1937 2197, GRID grid.169077.e, Department of Chemistry, , Purdue University, ; West Lafayette, IN 47907 USA
                [7 ]ISNI 0000 0004 0618 5819, GRID grid.418000.d, Present Address: Department of Plant Biology, , Carnegie Institution for Science, ; Stanford, CA 94305 USA
                Author information
                http://orcid.org/0000-0002-7100-1401
                http://orcid.org/0000-0002-8743-2847
                http://orcid.org/0000-0002-6360-7980
                http://orcid.org/0000-0003-0302-3470
                http://orcid.org/0000-0001-8774-4309
                http://orcid.org/0000-0001-6043-4132
                Article
                14477
                10.1038/s41467-020-14477-9
                6992735
                32001690
                d9cf3bd6-c0fb-4f46-b74a-e1929bfe2f09
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 19 August 2019
                : 8 January 2020
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100001809, National Natural Science Foundation of China (National Science Foundation of China);
                Award ID: 31771358
                Award Recipient :
                Categories
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                © The Author(s) 2020

                Uncategorized
                plant signalling,abiotic
                Uncategorized
                plant signalling, abiotic

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