Rodríguez Egea, Pedro Luís

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Now showing 1 - 10 of 62
  • Publication
    ESCRT-I Component VPS23A Affects ABA Signaling by Recognizing ABA Receptors for Endosomal Degradation
    (Oxford University Press (OUP), 2016-12-05) Yu, Feifei; Lou, Lijuan; Tian, Miaomiao; Li, Qingliang; Ding, Yanglin; Cao, Xiaoqiang; Wu, Yaorong; Belda Palazón, Borja; Rodríguez Egea, Pedro Luís; Yang, Shuhua; Xie, Qi; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; National Key Research and Development Program of China; National Natural Science Foundation of China
    [EN] Recent discovery of PYR/PYL/RCAR-type abscisic acid (ABA) receptors has become one of most significant advances in plant science in the past decade. In mammals, endosomal sorting acts as an important pathway to downregulate different types of receptors, but its role in plant hormone signaling is poorly understood. Here, we report that an ubiquitin E2-like protein, VPS23A, which is a key component of ESCRT-I, negatively regulates ABA signaling. VPS23A has epistatic relationship with PYR/PYL/RCAR-type ABA receptors and disruption of VPS23A enhanced the activity of key kinase OST1 in the ABA signaling pathway under ABA treatment. Moreover, VPS23A interacts with PYR1/PYLs and K63-linked diubiquitin, and PYL4 possesses K63-linked ubiquitinated modification in vivo. Further analysis revealed that VPS23A affects the subcellular localization of PYR1 and the stability of PYL4. Taken together, our results suggest that VPS23A affects PYR1/PYL4 via vacuole-mediated degradation, providing an advanced understanding of both the turnover of ABA receptors and ESCRTs in plant hormone signaling.
  • Publication
    Targeted degradation of abscisic acid receptors is mediated by the ubiquitin ligase substrate adaptor DDA1 in Arabidopsis
    (American Society of Plant Biologists, 2014-02) Irigoyen, María Luisa; Iniesto, Elisa; Rodríguez Solovey, Leisa Natacha; Puga, Maria Isabel; Yanagawa, Yuki; Pick, Elah; Strickland, Elizabeth; Paz-Ares, Javier; Wei, Ning; De Jaeger, Geert; Rodríguez Egea, Pedro Luís; Deng, Xing Wang; Rubio, Vicente; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; Ministerio de Economía y Competitividad; National Science Foundation, EEUU; National Institutes of Health, EEUU
    [EN] CULLIN4-RING E3 ubiquitin ligases (CRL4s) regulate key developmental and stress responses in eukaryotes. Studies in both animals and plants have led to the identification of many CRL4 targets as well as specific regulatory mechanisms that modulate their function. The latter involve COP10-DET1-DDB1 (CDD)-related complexes, which have been proposed to facilitate target recognition by CRL4, although the molecular basis for this activity remains largely unknown. Here, we provide evidence that Arabidopsis thaliana DET1-, DDB1-ASSOCIATED1 (DDA1), as part of the CDD complex, provides substrate specificity for CRL4 by interacting with ubiquitination targets. Thus, we show that DDA1 binds to the abscisic acid (ABA) receptor PYL8, as well as PYL4 and PYL9, in vivo and facilitates its proteasomal degradation. Accordingly, we found that DDA1 negatively regulates ABA-mediated developmental responses, including inhibition of seed germination, seedling establishment, and root growth. All other CDD components displayed a similar regulatory function, although they did not directly interact with PYL8. Interestingly, DDA1-mediated destabilization of PYL8 is counteracted by ABA, which protects PYL8 by limiting its polyubiquitination. Altogether, our data establish a function for DDA1 as a substrate receptor for CRL4-CDD complexes and uncover a mechanism for the desensitization of ABA signaling based on the regulation of ABA receptor stability.
  • Publication
    Structure-Based Modulation of the Ligand Sensitivity of a Tomato Dimeric Abscisic Acid Receptor Through a Glu to Asp Mutation in the Latch Loop
    (Frontiers Media SA, 2022-06-06) Infantes, Lourdes; Rivera-Moreno, Maria; Daniel-Mozo, Miguel; Benavente, Juan Luis; Ocaña-Cuesta, Javier; Coego, Alberto; Lozano Juste, Jorge; Rodríguez Egea, Pedro Luís; Albert, Armando; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; Agencia Estatal de Investigación; Ministerio de Ciencia e Innovación
    [EN] The binding of the plant phytohormone Abscisic acid (ABA) to the family of ABA receptors (PYR/PYL/RCAR) triggers plant responses to abiotic stress. Thus, the implementation of genetic or chemical strategies to modulate PYR/PYL activity might be biotechnologically relevant. We have employed the available structural information on the PYR/PYL receptors to design SlPYL1, a tomato receptor, harboring a single point mutation that displays enhanced ABA dependent and independent activity. Interestingly, crystallographic studies show that this mutation is not directly involved in ABA recognition or in the downstream phosphatase (PP2C) inhibitory interaction, rather, molecular dynamic based ensemble refinement restrained by crystallographic data indicates that it enhances the conformational variability required for receptor activation and it is involved in the stabilization of an active form of the receptor. Moreover, structural studies on this receptor have led to the identification of niacin as an ABA antagonist molecule in vivo. We have found that niacin blocks the ABA binding site by mimicking ABA receptor interactions, and the niacin interaction inhibits the biochemical activity of the receptor.
  • Publication
    Jasmonate signaling involves the abscisic acid receptor PYL4 to regulate metabolic reprogramming in Arabidopsis and tobacco
    (National Academy of Sciences, 2011-04-05) Lackman, Petri; González Guzmán, Miguel; Tilleman, Sofie; Carqueijeiro, Ines; Perez Cuellar, Amparo; Moses, Tessa; Seo, Mitsunori; Kanno, Yuri; Hakkinen, Suvi T; Van Montagu, Marc C. E; Thevelein, Johan; Maaheimo, Hannu; Oksman-Caldentey, Kirsi-Marja; Rodríguez Egea, Pedro Luís; Rischer, Heiko; Goossens, Alain; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; European Commission; Ministerio de Ciencia e Innovación
    [EN] The phytohormones jasmonates (JAs) constitute an important class of elicitors for many plant secondary metabolic pathways. However, JAs do not act independently but operate in complex networks with crosstalk to several other phytohormonal signaling pathways. Here, crosstalk was detected between the JA and abscisic acid (ABA) signaling pathways in the regulation of tobacco (Nicotiana tabacum) alkaloid biosynthesis. A tobacco gene from the PYR/PYL/RCAR family, NtPYL4, the expression of which is regulated by JAs, was found to encode a functional ABA receptor. NtPYL4 inhibited the type-2C protein phosphatases known to be key negative regulators of ABA signaling in an ABA-dependent manner. Overexpression of NtPYL4 in tobacco hairy roots caused a reprogramming of the cellular metabolism that resulted in a decreased alkaloid accumulation and conferred ABA sensitivity to the production of alkaloids. In contrast, the alkaloid biosynthetic pathway was not responsive to ABA in control tobacco roots. Functional analysis of the Arabidopsis (Arabidopsis thaliana) homologs of NtPYL4, PYL4 and PYL5, indicated that also in Arabidopsis altered PYL expression affected the JA response, both in terms of biomass and anthocyanin production. These findings define a connection between a component of the core ABA signaling pathway and the JA responses and contribute to the understanding of the role of JAs in balancing tradeoffs between growth and defense.
  • Publication
    Plant roots use a patterning mechanism to position lateral root branches toward available water
    (National Academy of Sciences, 2014-06-24) Bao, Yun; Aggarwal, Pooja; Robbins, Neil E. II; Sturrock, Craig J.; Thompson, Mark C.; Tan, Han Qi; Tham, Cliff; Duan, Lina; Rodríguez Egea, Pedro Luís; Vernoux, Teva; Mooney, Sacha J.; Bennet, Malcolm J.; Dinneny, Jose R.; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; Carnegie Institution; National Research Foundation, Singapur; National Science Foundation, EEUU; European Commission; Biotechnology and Biological Sciences Research Council, Reino Unido
    [EN] The architecture of the branched root system of plants is a major determinant of vigor. Water availability is known to impact root physiology and growth; however, the spatial scale at which this stimulus influences root architecture is poorly understood. Here we reveal that differences in the availability of water across the circumferential axis of the root create spatial cues that determine the position of lateral root branches. We show that roots of several plant species can distinguish between a wet surface and air environments and that this also impacts the patterning of root hairs, anthocyanins, and aerenchyma in a phenomenon we describe as hydropatterning. This environmental response is distinct from a touch response and requires available water to induce lateral roots along a contacted surface. X-ray microscale computed tomography and 3D reconstruction of soil-grown root systems demonstrate that such responses also occur under physiologically relevant conditions. Using early-stage lateral root markers, we show that hydropatterning acts before the initiation stage and likely determines the circumferential position at which lateral root founder cells are specified. Hydro-patterning is independent of endogenous abscisic acid signaling, distinguishing it from a classic water-stress response. Higher water availability induces the biosynthesis and transport of the lateral root-inductive signal auxin through local regulation of TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 and PIN-FORMED 3, both of which are necessary for normal hydropatterning. Our work suggests that water availability is sensed and interpreted at the suborgan level and locally patterns a wide variety of developmental processes in the root.
  • Publication
    The fungal sesquiterpenoid pyrenophoric acid B uses the plant ABA biosynthetic pathway to inhibit seed germination
    (Oxford University Press, 2019-10-01) Lozano Juste, Jorge; Masi, Marco; Cimmino, Alessio; Clement, Suzette; FERNÁNDEZ LÓPEZ, MARIA ANGELES; Antoni, Regina; Meyer, Susan; Rodríguez Egea, Pedro Luís; Evidente, Antonio; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; European Commission; Compagnia di San Paolo; U.S. Department of Agriculture; U.S. Department of the Interior; Ministerio de Educación, Cultura y Deporte; Università degli Studi di Napoli Federico II; Ministerio de Ciencia, Innovación y Universidades; Agencia Estatal de Investigación
    [EN] Pyrenophoric acid (P-Acid), P-Acid B, and P-Acid C are three phytotoxic sesquiterpenoids produced by the ascomycete seed pathogen Pyrenophora semeniperda, a fungus proposed as a mycoherbicide for biocontrol of cheatgrass, an extremely invasive weed. When tested in cheatgrass bioassays, these metabolites were able to delay seed germination, with P-Acid B being the most active compound. Here, we have investigated the cross-kingdom activity of P-Acid B and its mode of action, and found that it activates the abscisic acid (ABA) signaling pathway in order to inhibit seedling establishment. P-Acid B inhibits seedling establishment in wild-type Arabidopsis thaliana, while several mutants affected in the early perception as well as in downstream ABA signaling components were insensitive to the fungal compound. However, in spite of structural similarities between ABA and P-Acid B, the latter is not able to activate the PYR/PYL family of ABA receptors. Instead, we have found that P-Acid B uses the ABA biosynthesis pathway at the level of alcohol dehydrogenase ABA2 to reduce seedling establishment. We propose that the fungus P. semeniperda manipulates plant ABA biosynthesis as a strategy to reduce seed germination, increasing its ability to cause seed mortality and thereby increase its fitness through higher reproductive success.
  • Publication
    The Role of ABA in Plant Immunity is Mediated through the PYR1 Receptor
    (MDPI AG, 2020-08) García-Andrade, Javier; González, Beatriz; Gonzalez-Guzman, Miguel; Rodríguez Egea, Pedro Luís; Vera Vera, Pablo; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; Agencia Estatal de Investigación
    [EN] ABA is involved in plant responses to a broad range of pathogens and exhibits complex antagonistic and synergistic relationships with salicylic acid (SA) and ethylene (ET) signaling pathways, respectively. However, the specific receptor of ABA that triggers the positive and negative responses of ABA during immune responses remains unknown. Through a reverse genetic analysis, we identified that PYR1, a member of the family of PYR/PYL/RCAR ABA receptors, is transcriptionally upregulated and specifically perceives ABA during biotic stress, initiating downstream signaling mediated by ABA-activated SnRK2 protein kinases. This exerts a damping effect on SA-mediated signaling, required for resistance to biotrophic pathogens, and simultaneously a positive control over the resistance to necrotrophic pathogens controlled by ET. We demonstrated that PYR1-mediated signaling exerted control on a priori established hormonal cross-talk between SA and ET, thereby redirecting defense outputs. Defects in ABA/PYR1 signaling activated SA biosynthesis and sensitized plants for immune priming by poising SA-responsive genes for enhanced expression. As a trade-off effect,pyr1-mediated activation of the SA pathway blunted ET perception, which is pivotal for the activation of resistance towards fungal necrotrophs. The specific perception of ABA by PYR1 represented a regulatory node, modulating different outcomes in disease resistance.
  • Publication
    Root hydrotropism is controlled via a cortex-specific growth mechanism
    (Nature Publishing Group, 2017-06) Dietrich, Daniela; Pang, Lei; Kobayashi, Akie; Fozard, John A.; Boudolf, Veronique; Bhosale, Rahul; Nguyen, Tuan; Hiratsuka, Sotaro; Fujii, Nobuharu; Miyazawa, Yutaka; Bae, Tae-Woong; Wells, Darren M.; Owen, Markus R.; Band, Leah R.; Dyson, Rosemary J.; Rodríguez Egea, Pedro Luís; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; Belgian Federal Science Policy Office; Research Foundation Flanders; Ministry of Education, Culture, Sports, Science and Technology, Japón
    [EN] Plants can acclimate by using tropisms to link the direction of growth to environmental conditions. Hydrotropism allows roots to forage for water, a process known to depend on abscisic acid (ABA) but whose molecular and cellular basis remains unclear. Here we show that hydrotropism still occurs in roots after laser ablation removed the meristem and root cap. Additionally, targeted expression studies reveal that hydrotropism depends on the ABA signalling kinase SnRK2.2 and the hydrotropism-specific MIZ1, both acting specifically in elongation zone cortical cells. Conversely, hydrotropism, but not gravitropism, is inhibited by preventing differential cell-length increases in the cortex, but not in other cell types. We conclude that root tropic responses to gravity and water are driven by distinct tissue-based mechanisms. In addition, unlike its role in root gravitropism, the elongation zone performs a dual function during a hydrotropic response, both sensing a water potential gradient and subsequently undergoing differential growth.
  • Publication
    The PYL4 A194T mutant uncovers a key role of PYL4-PP2CA interaction for ABA signaling and plant drought resistance
    (American Society of Plant Biologists, 2013-09) Pizzio Bianchi, Gaston Alfredo; Rodriguez, Lesia; Antoni-Alandes, Regina; González Guzmán, Miguel; Yunta, Cristina; Merilo, Ebe; Kollist, Hannes; Albert, Armando; Rodríguez Egea, Pedro Luís; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas
    [EN] Because abscisic acid (ABA) is recognized as the critical hormonal regulator of plant stress physiology, elucidating its signaling pathway has raised promise for application in agriculture, for instance through genetic engineering of ABA receptors. PYRABACTIN RESISTANCE1/PYR1-LIKE (PYL)/REGULATORY COMPONENTS OF ABA RECEPTORS ABA receptors interact with high affinity and inhibit clade A phosphatases type-2C (PP2Cs) in an ABA-dependent manner. We generated an allele library composed of 10,000 mutant clones of Arabidopsis (Arabidopsis thaliana) PYL4 and selected mutations that promoted ABA-independent interaction with PP2CA/ABA-HYPERSENSITIVE3. In vitro protein-protein interaction assays and size exclusion chromatography confirmed that PYL4(A194T) was able to form stable complexes with PP2CA in the absence of ABA, in contrast to PYL4. This interaction did not lead to significant inhibition of PP2CA in the absence of ABA; however, it improved ABA-dependent inhibition of PP2CA. As a result, 35S: PYL4(A194T) plants showed enhanced sensitivity to ABA-mediated inhibition of germination and seedling establishment compared with 35S:PYL4 plants. Additionally, at basal endogenous ABA levels, whole-rosette gas exchange measurements revealed reduced stomatal conductance and enhanced water use efficiency compared with nontransformed or 35S:PYL4 plants and partial up-regulation of two ABA-responsive genes. Finally, 35S:PYL4(A194T) plants showed enhanced drought and dehydration resistance compared with nontransformed or 35S:PYL4 plants. Thus, we describe a novel approach to enhance plant drought resistance through allele library generation and engineering of a PYL4 mutation that enhances interaction with PP2CA.
  • Publication
    Depletion of abscisic acid levels in roots of flooded Carrizo citrange (Poncirus trifoliata L. Raf. x Citrus sinensis L. Osb.) plants is a stress-specific response associated to the differential expression of PYR/PYL/RCAR receptors
    (Springer-Verlag, 2017) Arbona, V.; Zandalinas, Sara I.; Manzi, M.; González Guzmán, Miguel; Rodríguez Egea, Pedro Luís; Gómez-Cadenas, Aurelio; Instituto Universitario Mixto de Biología Molecular y Celular de Plantas; Ministerio de Economía y Competitividad; Universitat Jaume I
    [EN] Soil flooding reduces root abscisic acid (ABA) levels in citrus, conversely to what happens under drought. Despite this reduction, microarray analyses suggested the existence of a residual ABA signaling in roots of flooded Carrizo citrange seedlings. The comparison of ABA metabolism and signaling in roots of flooded and water stressed plants of Carrizo citrange revealed that the hormone depletion was linked to the upregulation of CsAOG, involved in ABA glycosyl ester (ABAGE) synthesis, and to a moderate induction of catabolism (CsCYP707A, an ABA 8'-hydroxylase) and buildup of dehydrophaseic acid (DPA). Drought strongly induced both ABA biosynthesis and catabolism (CsNCED1, 9-cis-neoxanthin epoxycarotenoid dioxygenase 1, and CsCYP707A) rendering a significant hormone accumulation. In roots of flooded plants, restoration of control ABA levels after stress release was associated to the upregulation of CsBGLU18 (an ABA beta-glycosidase) that cleaves ABAGE. Transcriptional profile of ABA receptor genes revealed a different induction in response to soil flooding (CsPYL5) or drought (CsPYL8). These two receptor genes along with CsPYL1 were cloned and expressed in a heterologous system. Recombinant CsPYL5 inhibited Delta NHAB1 activity in vitro at lower ABA concentrations than CsPYL8 or CsPYL1, suggesting its better performance under soil flooding conditions. Both stress conditions induced ABA-responsive genes CsABI5 and CsDREB2A similarly, suggesting the occurrence of ABA signaling in roots of flooded citrus seedlings. The impact of reduced ABA levels in flooded roots on CsPYL5 expression along with its higher hormone affinity reinforce the role of this ABA receptor under soil-flooding conditions and explain the expression of certain ABA-responsive genes.