Emerging roles of protein kinase CK2 in abscisic acid signaling

The phytohormone abscisic acid (ABA) regulates many aspects of plant growth and development as well as responses to multiple stresses. Post-translational modifications such as phosphorylation or ubiquitination have pivotal roles in the regulation of ABA signaling. In addition to the positive regulator sucrose non-fermenting-1 related protein kinase 2 (SnRK2), the relevance of the role of other protein kinases, such as CK2, has been recently highlighted. We have recently established that CK2 phosphorylates the maize ortholog of open stomata 1 OST1, ZmOST1, suggesting a role of CK2 phosphorylation in the control of ZmOST1 protein degradation (Vilela et al., 2015). CK2 is a pleiotropic enzyme involved in multiple developmental and stress-responsive pathways. This review summarizes recent advances that taken together suggest a prominent role of protein kinase CK2 in ABA signaling and related processes

ZmSnRK2.8 responds to ABA through the SnRK2-PP2C complex

Understanding the responses of maize to abscisic acid (ABA) dependent drought tolerance is an important topic for the biotechnological application of functional mechanisms of stress adaptation. Key components that control and modulate stress adaptive pathways include SnRK2 (sucrose non-fermenting 1-related protein kinases 2) proteins. Recent studies indicate that SnRK2 are plant specific kinases that together with ABA ligand PYR/PYL/RCAR proteins and type 2C group A protein phosphatases constitute the central core of abscisic acid perception and signal transduction. Here, we study drought responses in maize by analyzing the mechanism of ZmSnRK2.8 in ABA signaling to establish relevant analogies with other plant species. ZmSnRK2.8 is a very closely related protein to Arabidopsis OST1 (open stomata 1) kinase with nuclear and cytosolic subcellular localization able to auto-phosphorylate S182 or T183 amino acids on its activation loop suggesting that phosphorylation at these sites may be a general mechanism for SnRK2 activation. In addition, ZmSnRK2.8 is activated by ABA and interacts with PP2C phosphatases in a constitutive, ABA independent manner. Together, our data suggest a conserved mechanism of plant responses to ABA and drought stress in maize and point to the potential use of this kinase in improving programs of drought tolerance in crops

The Heat Shock Factor A4A confers salt tolerance and is regulated by oxidative stress and the Mitogen-Activated Protein kinases, MPK3 and MPK6

Heat-shock factors (HSFs) are principal regulators of plant responses to several abiotic stresses. Here we show that estradiol-dependent induction of HSFA4A confers enhanced tolerance to salt and oxidative agents, whereas inactivation of HSFA4A results in hypersensitivity to salt stress in Arabidopsis. Estradiol-induction of HSFA4A in transgenic plants decreases, while the knockout hsfa4a mutation elevates hydrogen peroxide accumulation and lipid peroxidation. Overexpression of HSFA4A alters the transcription of a large set of genes regulated by oxidative stress. In yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays HSFA4A shows homomeric interaction which is reduced by alanine replacement of three conserved cysteine residues. HSFA4A interacts with mitogen-activated protein kinases MPK3 and MPK6 in yeast and plant cells. MPK3 and MPK6 phosphorylate HSFA4A in vitro on three distinct sites, Ser309 being the major phosphorylation site. Activation of the MPK3 and MPK6 MAPK pathway led to the transcriptional activation of the heat-shock protein gene HSP17.6A. In agreement that mutation of Ser309 to alanine strongly diminished phosphorylation of HSFA4A, it also strongly reduced the transcriptional activation of HSP17.6A. These data suggest that HSFA4A is a substrate of the MPK3/6 signalling and it regulates stress responses in Arabidopsis

Functional and molecular characterization of maize open stomata 1 protein kinase

[eng] Plant growth and productivity are compromised by environmental stresses such as pathogens, extreme temperatures, drought and high salinity. Being sessile organisms, plants had to develop different physiologic and biochemical strategies to cope with these potential harmful situations. Drought in particular is one of the major environmental stresses that plants are forced to endure during their life cycle. The adaptation to water deficit is controlled by a cascade of molecular networks that start with the perception of water shortage which leads to increases in the ABA levels. Even though the ABA signalling model is well described for Arabidopsis, little is known for other plant species. With this thesis we proposed to increase the knowledge of maize response to drought, focusing on a maize kinase of the SnRK2 family ‐ ZmSnRK2.8/ZmOST1 ‐ which is highly homologous to the Arabidopsis OST1. We divided our work on three chapters, namely the biochemical characterization of ZmOST1, the functional characterization of ZmOST1 and the study of ZmOST1 regulation. 1) ZmOST1 biochemical characterization With Chapter 1 we characterized ZmOST1 at the biochemical level, making parallels with the Arabidopsis system whenever pertinent. We found a very close biochemical relationship between the maize and Arabidopsis kinases that suggests a conserved mechanism of plant responses to ABA and drought stress and point to the potential use of this kinase in improvement programs of drought tolerance in crops. 2) ZmOST1 functional characterization With Chapter 2 we described ZmOST1 as a functional kinase that is activated by different osmotic stresses and that is able to complement the Arabidopsis ost1‐2 mutant with effects on stomata closure. We also present a transcription factor of the NAC superfamily (ZmSNAC1) as a novel cognate substrate of ZmOST1. Under abiotic stresses ZmOST1 is capable of phosphorylating this transcription factor with further implications on stomata regulation. 3) ZmOST1 regulation With the results presented in Chapter 3 a larger picture of ABA signalling appears that implicates new partners on ZmOST1 regulation, specifically the CK2 kinase and the proteasome degradation. Ample evidence is shown suggesting CK2 phosphorylation is implicated in ABA signalling by affecting ZmOST1 localization, protein levels, protein degradation and interaction with PP2C phosphatases. At the plant level, overexpressing ZmOST1 mutagenized on the CK2 loci of phosphorylation grants several potential beneficial traits that could prove important for crop biotechnology, such as higher protein levels, better protein stability, enhanced phosphorylation activity and better stomata regulation. Working model: Taken the results presented in this thesis together, we propose a change in the current ABA signalling model. First we believe that there is an important role for CK2 in ABA sensing and SnRK2 activation that could affect the binding of the kinase to the PP2C phosphatises and regulate SnRK2 through degradation. Second, we propose that, apart from the always off and transiently fast on/off modes of SnRK2 activity, there is a third always on mechanism in which the kinase is fully detached from the phosphatase.[spa] La sequía es actualmente el factor abiótico más limitante para el crecimiento de las plantas y se está agravando con los actuales cambios climáticos, aumento de población y reducción de las reservas de agua. Se estima que en el 2050 el 50% de las tierras cultivadas tengan problemas de salinidad o sequía. Por ello, la intensificación de la agricultura y el desarrollo de la mejora biotecnológica de adaptación al estrés son áreas que tienen que reforzarse. En esta tesis se pretende ampliar los conocimientos sobre la respuesta del maíz a la sequía haciendo un estudio profundizado de una quinasa de tipo SnRK2, designada ZmOST1 que está implicada en la respuesta de las plantas al déficit hídrico. - Capítulo 1: Caracterización bioquímica de la quinasa de maíz ZmOST1 en que se establece que ZmOST1 se localiza en el núcleo y citoplasma, se activa por ABA, es capaz de autofosforilar su centro activo y directamente interacciona con una fosfatasa ZmPP2C a través de su dominio regulador. - Capítulo 2: Caracterización fisiológica de ZmOST1. Se determinan los niveles de expresión y de actividad de la quinasa en diferentes tratamientos y estadios de desarrollo; se establece que ZmOST1 es capaz de recuperar el fenotipo de cierre estomático en mutantes OST1 de Arabidopsis; y se identifica un factor de transcripción que se caracteriza como un nuevo substrato de esta quinasa. - Capítulo 3: Se describe una nueva ruta de regulación de ZmOST1 en que esta quinasa es fosforilada por la CK2 (casein kinase 2) en el dominio regulador. Mutagenizando los residuos diana de la CK2 en la ZmOST1 lleva a una mayor acumulación, una menor degradación por el proteasoma y una hipersensibilidad a ABA

Emerging roles of protein kinase CK2 in abscisic acid signaling

The phytohormone abscisic acid (ABA) regulates many aspects of plant growth and development as well as responses to multiple stresses. Post-translational modifications such as phosphorylation or ubiquitination have pivotal roles in the regulation of ABA signaling. In addition to the positive regulator sucrose non-fermenting-1 related protein kinase 2 (SnRK2), the relevance of the role of other protein kinases, such as CK2, has been recently highlighted. We have recently established that CK2 phosphorylates the maize ortholog of open stomata 1 OST1, ZmOST1, suggesting a role of CK2 phosphorylation in the control of ZmOST1 protein degradation (Vilela et al., 2015). CK2 is a pleiotropic enzyme involved in multiple developmental and stress-responsive pathways. This review summarizes recent advances that taken together suggest a prominent role of protein kinase CK2 in ABA signaling and related processes.This work was supported by grant BIO2012-31860 (MICINN, Spain) to MP.Peer reviewedPeer Reviewe

Emerging roles of protein kinase CK2 in abscisic acid signaling

The phytohormone abscisic acid (ABA) regulates many aspects of plant growth and development as well as responses to multiple stresses. Post-translational modifications such as phosphorylation or ubiquitination have pivotal roles in the regulation of ABA signaling. In addition to the positive regulator sucrose non-fermenting-1 related protein kinase 2 (SnRK2), the relevance of the role of other protein kinases, such as CK2, has been recently highlighted. We have recently established that CK2 phosphorylates the maize ortholog of open stomata 1 OST1, ZmOST1, suggesting a role of CK2 phosphorylation in the control of ZmOST1 protein degradation (Vilela et al., 2015). CK2 is a pleiotropic enzyme involved in multiple developmental and stress-responsive pathways. This review summarizes recent advances that taken together suggest a prominent role of protein kinase CK2 in ABA signaling and related processes

ZmSnRK2.8 is involved in stomatal closure and phosphorylates two maize transcription factors

Trabajo presentado al 20th International Plant Growth Substances Association (IPGSA) Conference, celebrada en 2010.Peer reviewe

Drought tolerance in Zea mays: ZmOST1 characterization and regulation its potential substrate ZmbHLH transcription factor

Resumen del trabajo presentado al Icrea Workshop: From model systems to crops, challenges for a new era in plant biology, celebrado del 7 al 8 de mayo de 2014 en Barcelona.Peer reviewe