Home LiteratureArticle Details
PMID: 28438792 Published · ppublish English Journal Article

Reconstitution of Abscisic Acid Signaling from the Receptor to DNA via bHLH Transcription Factors.

Plant physiology ·Vol. 174 ·No. 2 ·2017-06-00 ·Pages 815-822

Takahashi Y, Ebisu Y, Shimazaki KI

Abstract

The plant hormone abscisic acid (ABA) confers drought tolerance in plants through stomatal closure and regulation of gene expression. The complex consisting of the ABA receptor PYRABACTIN RESISTANCE/REGULATORY COMPONENTS OF ABA RECEPTOR (PYR/RCAR), type 2C protein phosphatase (PP2C), and SNF1-related protein kinase 2 (SnRK2) has a key role in ABA signaling. Basic helix-loop-helix (bHLH) transcriptional activator ABA-RESPONSIVE KINASE SUBSTRATE1 (AKS1, also known as FBH3) is released from DNA by phosphorylation-induced monomerization in response to ABA in guard cells. Here we reconstituted the release of AKS1 from DNA via the ABA signaling core complex in vitro. We first obtained evidence to confirm that AKS1 is an endogenous substrate for SnRK2s. Phosphorylation of AKS1 and activation of SnRK2 showed the same time course in response to ABA in guard cells. AKS1 was bound to SnRK2.6 in vivo. Three ABA-responsive SnRK2s (SnRK2.2/SRK2D, SnRK2.3/SRK2I, and SnRK2.6/SRK2E/OST1) phosphorylated AKS1 in vitro, and the phosphorylation was eliminated by the triple mutation of SnRK2s in plants. We reconstituted the AKS1 phosphorylation in vitro via the signaling complex containing the ABA receptor PYR1, a PP2C, HYPERSENSITIVE TO ABA1 (HAB1), and a protein kinase, SnRK2.6 in response to ABA We further reconstituted the release of AKS1 from the target gene of POTASSIUM CHANNEL IN ARABIDOPSIS THALIANA 1 (KAT1) via the complex in response to ABA These results demonstrate that AKS1 provides a link between the signaling complex and ABA-responsive genes and furnish evidence for a minimal signaling mechanism from ABA perception to DNA.

MeSH Terms
Abscisic Acid/metabolism Arabidopsis/genetics,metabolism Arabidopsis Proteins/genetics,metabolism Basic Helix-Loop-Helix Transcription Factors/genetics,metabolism DNA, Plant/metabolism Membrane Transport Proteins/genetics,metabolism Phosphoprotein Phosphatases/genetics,metabolism Phosphorylation Plant Cells/metabolism Plants, Genetically Modified Potassium Channels, Inwardly Rectifying/genetics,metabolism Promoter Regions, Genetic Protein Kinases/genetics,metabolism Protein Serine-Threonine Kinases/genetics,metabolism Signal Transduction
Chemicals
Arabidopsis Proteins Basic Helix-Loop-Helix Transcription Factors DNA, Plant FBH3 protein, Arabidopsis KAT1 protein, Arabidopsis Membrane Transport Proteins Potassium Channels, Inwardly Rectifying Pyr1 protein, Arabidopsis SnRK2 protein, Arabidopsis Abscisic Acid Protein Kinases OST1 protein, Arabidopsis Protein Serine-Threonine Kinases HAB1 protein, Arabidopsis Phosphoprotein Phosphatases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Takahashi Yohei ORCID
Department of Biology, Faculty of Science, Kyushu University 744 Motooka, Fukuoka 819-0395, Japan yotakahashi@ucsd.edu.
Ebisu Yuta
Department of Biology, Faculty of Science, Kyushu University 744 Motooka, Fukuoka 819-0395, Japan.
Shimazaki Ken-Ichiro ORCID
Department of Biology, Faculty of Science, Kyushu University 744 Motooka, Fukuoka 819-0395, Japan yotakahashi@ucsd.edu.
References (51)
51 references, click to expand
  1. Immunopurification and Mass Spectrometry Identifies Protein Phosphatase 2A (PP2A) and BIN2/GSK3 as Regulators of AKS Transcription Factors in Arabidopsis.
    Mol Plant. 2017 Feb 13;10 (2):345-348 PMID: 27769939
  2. Selective mobility and sensitivity to SNAREs is exhibited by the Arabidopsis KAT1 K+ channel at the plasma membrane.
    Plant Cell. 2006 Apr;18(4):935-54 PMID: 16531497
  3. Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL family of START proteins.
    Science. 2009 May 22;324(5930):1068-71 PMID: 19407142
  4. Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis.
    Nat Protoc. 2007;2(7):1565-72 PMID: 17585298
  5. Genetics and phosphoproteomics reveal a protein phosphorylation network in the abscisic acid signaling pathway in Arabidopsis thaliana.
    Sci Signal. 2013 Apr 09;6(270):rs8 PMID: 23572148
  6. Inhibition of the Arabidopsis bHLH transcription factor by monomerization through abscisic acid-induced phosphorylation.
    Plant J. 2016 Sep;87(6):559-67 PMID: 27227462
  7. Structural basis of abscisic acid signalling.
    Nature. 2009 Dec 3;462(7273):609-14 PMID: 19855379
  8. Arabidopsis mutant deficient in 3 abscisic acid-activated protein kinases reveals critical roles in growth, reproduction, and stress.
    Proc Natl Acad Sci U S A. 2009 May 19;106(20):8380-5 PMID: 19420218
  9. Biochemical characterization of plasma membrane H+-ATPase activation in guard cell protoplasts of Arabidopsis thaliana in response to blue light.
    Plant Cell Physiol. 2005 Jun;46(6):955-63 PMID: 15821287
  10. In vitro reconstitution of an abscisic acid signalling pathway.
    Nature. 2009 Dec 3;462(7273):660-4 PMID: 19924127
  11. Monitoring the expression pattern of around 7,000 Arabidopsis genes under ABA treatments using a full-length cDNA microarray.
    Funct Integr Genomics. 2002 Nov;2(6):282-91 PMID: 12444421
  12. Three SnRK2 protein kinases are the main positive regulators of abscisic acid signaling in response to water stress in Arabidopsis.
    Plant Cell Physiol. 2009 Dec;50(12 ):2123-32 PMID: 19880399
  13. Regulators of PP2C phosphatase activity function as abscisic acid sensors.
    Science. 2009 May 22;324(5930):1064-8 PMID: 19407143
  14. Arabidopsis nanodomain-delimited ABA signaling pathway regulates the anion channel SLAH3.
    Proc Natl Acad Sci U S A. 2013 May 14;110(20):8296-301 PMID: 23630285
  15. The regulatory domain of SRK2E/OST1/SnRK2.6 interacts with ABI1 and integrates abscisic acid (ABA) and osmotic stress signals controlling stomatal closure in Arabidopsis.
    J Biol Chem. 2006 Feb 24;281(8):5310-8 PMID: 16365038
  16. A gate-latch-lock mechanism for hormone signalling by abscisic acid receptors.
    Nature. 2009 Dec 3;462(7273):602-8 PMID: 19898420
  17. Molecular basis of the core regulatory network in ABA responses: sensing, signaling and transport.
    Plant Cell Physiol. 2010 Nov;51(11):1821-39 PMID: 20980270
  18. Reconstitution of abscisic acid activation of SLAC1 anion channel by CPK6 and OST1 kinases and branched ABI1 PP2C phosphatase action.
    Proc Natl Acad Sci U S A. 2012 Jun 26;109(26):10593-8 PMID: 22689970
  19. Protein phosphatases 2C regulate the activation of the Snf1-related kinase OST1 by abscisic acid in Arabidopsis.
    Plant Cell. 2009 Oct;21(10 ):3170-84 PMID: 19855047
  20. The trafficking protein SYP121 of Arabidopsis connects programmed stomatal closure and K⁺ channel activity with vegetative growth.
    Plant J. 2012 Jan;69(2):241-51 PMID: 21914010
  21. Physiological and transcriptional memory in guard cells during repetitive dehydration stress.
    New Phytol. 2015 Jan;205(2):596-607 PMID: 25345749
  22. Type 2C protein phosphatases directly regulate abscisic acid-activated protein kinases in Arabidopsis.
    Proc Natl Acad Sci U S A. 2009 Oct 13;106(41):17588-93 PMID: 19805022
  23. Common and unique elements of the ABA-regulated transcriptome of Arabidopsis guard cells.
    BMC Genomics. 2011 May 09;12:216 PMID: 21554708
  24. Activity of guard cell anion channel SLAC1 is controlled by drought-stress signaling kinase-phosphatase pair.
    Proc Natl Acad Sci U S A. 2009 Dec 15;106(50):21425-30 PMID: 19955405
  25. In planta changes in protein phosphorylation induced by the plant hormone abscisic acid.
    Proc Natl Acad Sci U S A. 2010 Sep 7;107(36):15986-91 PMID: 20733066
  26. Guard cell signal transduction network: advances in understanding abscisic acid, CO2, and Ca2+ signaling.
    Annu Rev Plant Biol. 2010;61:561-91 PMID: 20192751
  27. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses.
    Annu Rev Plant Biol. 2006;57:781-803 PMID: 16669782
  28. Mechanisms of abscisic acid-mediated control of stomatal aperture.
    Curr Opin Plant Biol. 2015 Dec;28:154-62 PMID: 26599955
  29. A protein kinase-phosphatase pair interacts with an ion channel to regulate ABA signaling in plant guard cells.
    Proc Natl Acad Sci U S A. 2009 Dec 15;106(50):21419-24 PMID: 19955427
  30. Abscisic Acid synthesis and response.
    Arabidopsis Book. 2013 Nov 01;11:e0166 PMID: 24273463
  31. Protein phosphorylation and binding of a 14-3-3 protein in Vicia guard cells in response to ABA.
    Plant Cell Physiol. 2007 Aug;48(8):1182-91 PMID: 17634179
  32. Structural mechanism of abscisic acid binding and signaling by dimeric PYR1.
    Science. 2009 Dec 4;326(5958):1373-9 PMID: 19933100
  33. Pivotal role of the AREB/ABF-SnRK2 pathway in ABRE-mediated transcription in response to osmotic stress in plants.
    Physiol Plant. 2013 Jan;147(1):15-27 PMID: 22519646
  34. The Arabidopsis ABA-activated kinase OST1 phosphorylates the bZIP transcription factor ABF3 and creates a 14-3-3 binding site involved in its turnover.
    PLoS One. 2010 Nov 10;5(11):e13935 PMID: 21085673
  35. Calcium specificity signaling mechanisms in abscisic acid signal transduction in Arabidopsis guard cells.
    Elife. 2015 Jul 20;4:null PMID: 26192964
  36. The basic helix-loop-helix transcription factor family in plants: a genome-wide study of protein structure and functional diversity.
    Mol Biol Evol. 2003 May;20(5):735-47 PMID: 12679534
  37. The Arabidopsis basic/helix-loop-helix transcription factor family.
    Plant Cell. 2003 Aug;15(8):1749-70 PMID: 12897250
  38. ABA perception and signalling.
    Trends Plant Sci. 2010 Jul;15(7):395-401 PMID: 20493758
  39. Gain-of-function and loss-of-function phenotypes of the protein phosphatase 2C HAB1 reveal its role as a negative regulator of abscisic acid signalling.
    Plant J. 2004 Feb;37(3):354-69 PMID: 14731256
  40. Four Arabidopsis AREB/ABF transcription factors function predominantly in gene expression downstream of SnRK2 kinases in abscisic acid signalling in response to osmotic stress.
    Plant Cell Environ. 2015 Jan;38(1):35-49 PMID: 24738645
  41. bHLH transcription factors that facilitate K⁺ uptake during stomatal opening are repressed by abscisic acid through phosphorylation.
    Sci Signal. 2013 Jun 18;6(280):ra48 PMID: 23779086
  42. AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis.
    Plant Cell. 2005 Dec;17(12):3470-88 PMID: 16284313
  43. AREB1, AREB2, and ABF3 are master transcription factors that cooperatively regulate ABRE-dependent ABA signaling involved in drought stress tolerance and require ABA for full activation.
    Plant J. 2010 Feb;61(4):672-85 PMID: 19947981
  44. Abscisic acid triggers the endocytosis of the arabidopsis KAT1 K+ channel and its recycling to the plasma membrane.
    Curr Biol. 2007 Aug 21;17(16):1396-402 PMID: 17683934
  45. Quantitative phosphoproteomics identifies SnRK2 protein kinase substrates and reveals the effectors of abscisic acid action.
    Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):11205-10 PMID: 23776212
  46. Microarray expression analyses of Arabidopsis guard cells and isolation of a recessive abscisic acid hypersensitive protein phosphatase 2C mutant.
    Plant Cell. 2004 Mar;16(3):596-615 PMID: 14973164
  47. A transcription factor hierarchy defines an environmental stress response network.
    Science. 2016 Nov 4;354(6312): PMID: 27811239
  48. Abscisic acid: emergence of a core signaling network.
    Annu Rev Plant Biol. 2010;61:651-79 PMID: 20192755
  49. Three Arabidopsis SnRK2 protein kinases, SRK2D/SnRK2.2, SRK2E/SnRK2.6/OST1 and SRK2I/SnRK2.3, involved in ABA signaling are essential for the control of seed development and dormancy.
    Plant Cell Physiol. 2009 Jul;50(7):1345-63 PMID: 19541597
  50. Abscisic acid-dependent multisite phosphorylation regulates the activity of a transcription activator AREB1.
    Proc Natl Acad Sci U S A. 2006 Feb 7;103(6):1988-93 PMID: 16446457
  51. C2-domain abscisic acid-related proteins mediate the interaction of PYR/PYL/RCAR abscisic acid receptors with the plasma membrane and regulate abscisic acid sensitivity in Arabidopsis.
    Plant Cell. 2014 Dec;26(12):4802-20 PMID: 25465408
Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
1532-2548
DOI
10.1104/pp.16.01825
Email
yotakahashi@ucsd.edu
Published
2017-06-00
Epub
2017-00-24
Pages
815-822
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC5462015
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.16.01825 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2