In vitro reconstitution of an abscisic acid signalling pathway

Handle URI:
http://hdl.handle.net/10754/325269
Title:
In vitro reconstitution of an abscisic acid signalling pathway
Authors:
Fujii, Hiroaki ( 0000-0002-0013-5891 ) ; Chinnusamy, Viswanathan; Rodrigues, Americo; Rubio, Silvia; Antoni, Regina; Park, Sang-Youl; Cutler, Sean R.; Sheen, Jen; Rodriguez, Pedro L.; Zhu, Jian-Kang ( 0000-0001-5134-731X )
Abstract:
The phytohormone abscisic acid (ABA) regulates the expression of many genes in plants; it has critical functions in stress resistance and in growth and development. Several proteins have been reported to function as ABA receptors, and many more are known to be involved in ABA signalling. However, the identities of ABA receptors remain controversial and the mechanism of signalling from perception to downstream gene expression is unclear. Here we show that by combining the recently identified ABA receptor PYR1 with the type 2C protein phosphatase (PP2C) ABI1, the serine/threonine protein kinase SnRK2.6/OST1 and the transcription factor ABF2/AREB1, we can reconstitute ABA-triggered phosphorylation of the transcription factor in vitro. Introduction of these four components into plant protoplasts results in ABA-responsive gene expression. Protoplast and test-tube reconstitution assays were used to test the function of various members of the receptor, protein phosphatase and kinase families. Our results suggest that the default state of the SnRK2 kinases is an autophosphorylated, active state and that the SnRK2 kinases are kept inactive by the PP2Cs through physical interaction and dephosphorylation. We found that in the presence of ABA, the PYR/PYL (pyrabactin resistance 1/PYR1-like) receptor proteins can disrupt the interaction between the SnRK2s and PP2Cs, thus preventing the PP2C-mediated dephosphorylation of the SnRK2s and resulting in the activation of the SnRK2 kinases. Our results reveal new insights into ABA signalling mechanisms and define a minimal set of core components of a complete major ABA signalling pathway. © 2009 Macmillan Publishers Limited. All rights reserved.
KAUST Department:
Plant Stress Genomics Research Lab
Citation:
Fujii H, Chinnusamy V, Rodrigues A, Rubio S, Antoni R, et al. (2009) In vitro reconstitution of an abscisic acid signalling pathway. Nature 462: 660-664. doi:10.1038/nature08599.
Publisher:
Nature Publishing Group
Journal:
Nature
Issue Date:
18-Nov-2009
DOI:
10.1038/nature08599
PubMed ID:
19924127
PubMed Central ID:
PMC2803041
Type:
Article
ISSN:
00280836
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorFujii, Hiroakien
dc.contributor.authorChinnusamy, Viswanathanen
dc.contributor.authorRodrigues, Americoen
dc.contributor.authorRubio, Silviaen
dc.contributor.authorAntoni, Reginaen
dc.contributor.authorPark, Sang-Youlen
dc.contributor.authorCutler, Sean R.en
dc.contributor.authorSheen, Jenen
dc.contributor.authorRodriguez, Pedro L.en
dc.contributor.authorZhu, Jian-Kangen
dc.date.accessioned2014-08-27T09:43:49Zen
dc.date.available2014-08-27T09:43:49Zen
dc.date.issued2009-11-18en
dc.identifier.citationFujii H, Chinnusamy V, Rodrigues A, Rubio S, Antoni R, et al. (2009) In vitro reconstitution of an abscisic acid signalling pathway. Nature 462: 660-664. doi:10.1038/nature08599.en
dc.identifier.issn00280836en
dc.identifier.pmid19924127en
dc.identifier.doi10.1038/nature08599en
dc.identifier.urihttp://hdl.handle.net/10754/325269en
dc.description.abstractThe phytohormone abscisic acid (ABA) regulates the expression of many genes in plants; it has critical functions in stress resistance and in growth and development. Several proteins have been reported to function as ABA receptors, and many more are known to be involved in ABA signalling. However, the identities of ABA receptors remain controversial and the mechanism of signalling from perception to downstream gene expression is unclear. Here we show that by combining the recently identified ABA receptor PYR1 with the type 2C protein phosphatase (PP2C) ABI1, the serine/threonine protein kinase SnRK2.6/OST1 and the transcription factor ABF2/AREB1, we can reconstitute ABA-triggered phosphorylation of the transcription factor in vitro. Introduction of these four components into plant protoplasts results in ABA-responsive gene expression. Protoplast and test-tube reconstitution assays were used to test the function of various members of the receptor, protein phosphatase and kinase families. Our results suggest that the default state of the SnRK2 kinases is an autophosphorylated, active state and that the SnRK2 kinases are kept inactive by the PP2Cs through physical interaction and dephosphorylation. We found that in the presence of ABA, the PYR/PYL (pyrabactin resistance 1/PYR1-like) receptor proteins can disrupt the interaction between the SnRK2s and PP2Cs, thus preventing the PP2C-mediated dephosphorylation of the SnRK2s and resulting in the activation of the SnRK2 kinases. Our results reveal new insights into ABA signalling mechanisms and define a minimal set of core components of a complete major ABA signalling pathway. © 2009 Macmillan Publishers Limited. All rights reserved.en
dc.language.isoenen
dc.publisherNature Publishing Groupen
dc.rightsUsers may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#termsen
dc.rightsArchived with thanks to Natureen
dc.subjectabscisic aciden
dc.subjectphosphoprotein phosphataseen
dc.subjectphosphoprotein phosphatase 2en
dc.subjectprotein serine threonine kinaseen
dc.subjecttranscription factoren
dc.subjectabscisic aciden
dc.subjectbioassayen
dc.subjectenzyme activityen
dc.subjectgene expressionen
dc.subjectproteinen
dc.subjectsignalingen
dc.subjectgene expressionen
dc.subjectin vitro studyen
dc.subjectplant geneen
dc.subjectprotein dephosphorylationen
dc.subjectprotein interactionen
dc.subjectprotein phosphorylationen
dc.subjectprotoplasten
dc.subjectstressen
dc.subjectAbscisic Aciden
dc.subjectArabidopsisen
dc.subjectArabidopsis Proteinsen
dc.subjectGene Expression Regulation, Planten
dc.subjectMutationen
dc.subjectPhenotypeen
dc.subjectPhosphorylationen
dc.subjectProtoplastsen
dc.subjectSignal Transductionen
dc.subjectStress, Physiologicalen
dc.titleIn vitro reconstitution of an abscisic acid signalling pathwayen
dc.typeArticleen
dc.contributor.departmentPlant Stress Genomics Research Laben
dc.identifier.journalNatureen
dc.identifier.pmcidPMC2803041en
dc.eprint.versionPost-printen
dc.contributor.institutionDepartment of Botany and Plant Sciences, University of California at Riverside, Riverside, CA 92521, United Statesen
dc.contributor.institutionInstituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-UPV, ES-46022 Valencia, Spainen
dc.contributor.institutionDepartment of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, United Statesen
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
kaust.authorFujii, Hiroakien
kaust.authorChinnusamy, Viswanathanen
kaust.authorZhu, Jian-Kangen

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