bHLH003, bHLH013 and bHLH017 Are New Targets of JAZ Repressors Negatively Regulating JA Responses

Cell reprogramming in response to jasmonates requires a tight control of transcription that is achieved by the activity of JA-related transcription factors (TFs). Among them, MYC2, MYC3 and MYC4 have been described as activators of JA responses. Here we characterized the function of bHLH003, bHLH013 and bHLH017 that conform a phylogenetic clade closely related to MYC2, MYC3 and MYC4. We found that these bHLHs form homo- and heterodimers and also interact with JAZ repressors in vitro and in vivo. Phenotypic analysis of JA-regulated processes, including root and rosette growth, anthocyanin accumulation, chlorophyll loss and resistance to Pseudomonas syringae, on mutants and overexpression lines, suggested that these bHLHs are repressors of JA responses. bHLH003, bHLH013 and bHLH017 are mainly nuclear proteins and bind DNA with similar specificity to that of MYC2, MYC3 and MYC4, but lack a conserved activation domain, suggesting that repression is achieved by competition for the same cis-regulatory elements. Moreover, expression of bHLH017 is induced by JA and depends on MYC2, suggesting a negative feed-back regulation of the activity of positive JA-related TFs. Our results suggest that the competition between positive and negative TFs determines the output of JA-dependent transcriptional activation.

[1]  S. Stolz,et al.  Role of NINJA in root jasmonate signaling , 2013, Proceedings of the National Academy of Sciences.

[2]  Hongwei Guo,et al.  The bHLH Subgroup IIId Factors Negatively Regulate Jasmonate-Mediated Plant Defense and Development , 2013, PLoS genetics.

[3]  R. Solano,et al.  JASMONATE-INSENSITIVE1 Encodes a MYC Transcription Factor Essential to Discriminate between Different Jasmonate-Regulated Defense Responses in Arabidopsis , 2004, The Plant Cell Online.

[4]  Martin Hammarström,et al.  Rapid screening for improved solubility of small human proteins produced as fusion proteins in Escherichia coli , 2002, Protein science : a publication of the Protein Society.

[5]  S. Reinbothe,et al.  Plant oxylipins: role of jasmonic acid during programmed cell death, defence and leaf senescence , 2009, The FEBS journal.

[6]  D. Inzé,et al.  Mapping methyl jasmonate-mediated transcriptional reprogramming of metabolism and cell cycle progression in cultured Arabidopsis cells , 2008, Proceedings of the National Academy of Sciences.

[7]  G. Howe,et al.  Cytochrome P450 CYP94B3 mediates catabolism and inactivation of the plant hormone jasmonoyl-L-isoleucine , 2011, Proceedings of the National Academy of Sciences.

[8]  Martin Kuiper,et al.  Targeted interactomics reveals a complex core cell cycle machinery in Arabidopsis thaliana , 2010, Molecular systems biology.

[9]  R. Solano,et al.  The jasmonate pathway: the ligand, the receptor and the core signalling module. , 2009, Current opinion in plant biology.

[10]  W. Peng,et al.  The Jasmonate-ZIM-Domain Proteins Interact with the WD-Repeat/bHLH/MYB Complexes to Regulate Jasmonate-Mediated Anthocyanin Accumulation and Trichome Initiation in Arabidopsis thaliana[C][W] , 2011, Plant Cell.

[11]  D. Inzé,et al.  NINJA connects the co-repressor TOPLESS to jasmonate signalling , 2010, Nature.

[12]  Mathew G. Lewsey,et al.  Arabidopsis Basic Helix-Loop-Helix Transcription Factors MYC2, MYC3, and MYC4 Regulate Glucosinolate Biosynthesis, Insect Performance, and Feeding Behavior[W][OPEN] , 2013, Plant Cell.

[13]  S. Prat,et al.  Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis. , 2004, Genes & development.

[14]  H. Ohta,et al.  Basic Helix-Loop-Helix Transcription Factors JASMONATE-ASSOCIATED MYC2-LIKE1 (JAM1), JAM2, and JAM3 Are Negative Regulators of Jasmonate Responses in Arabidopsis1[W][OPEN] , 2013, Plant Physiology.

[15]  Wei Li,et al.  Derepression of ethylene-stabilized transcription factors (EIN3/EIL1) mediates jasmonate and ethylene signaling synergy in Arabidopsis , 2011, Proceedings of the National Academy of Sciences.

[16]  M. Pagni,et al.  A Downstream Mediator in the Growth Repression Limb of the Jasmonate Pathway[W][OA] , 2007, The Plant Cell Online.

[17]  C. Wasternack,et al.  Biochemical and Molecular Characterization of a Hydroxyjasmonate Sulfotransferase from Arabidopsis thaliana * , 2003, The Journal of Biological Chemistry.

[18]  Jonathan D. G. Jones,et al.  Evidence for Network Evolution in an Arabidopsis Interactome Map , 2011, Science.

[19]  M. Links,et al.  Genome-Wide Analysis of Ethylene-Responsive Element Binding Factor-Associated Amphiphilic Repression Motif-Containing Transcriptional Regulators in Arabidopsis1[W][OA] , 2010, Plant Physiology.

[20]  Hur-Song Chang,et al.  Expression profiling reveals COI1 to be a key regulator of genes involved in wound- and methyl jasmonate-induced secondary metabolism, defence, and hormone interactions , 2005, Plant Molecular Biology.

[21]  R. Solano,et al.  Pull-down analysis of interactions among jasmonic acid core signaling proteins. , 2013, Methods in molecular biology.

[22]  J. Micol,et al.  The JAZ family of repressors is the missing link in jasmonate signalling , 2007, Nature.

[23]  G. Howe,et al.  JAZ8 Lacks a Canonical Degron and Has an EAR Motif That Mediates Transcriptional Repression of Jasmonate Responses in Arabidopsis[C][W] , 2012, Plant Cell.

[24]  Anthony L. Schilmiller,et al.  COI1 is a critical component of a receptor for jasmonate and the bacterial virulence factor coronatine , 2008, Proceedings of the National Academy of Sciences.

[25]  P. Figueroa,et al.  Characterization of JAZ-interacting bHLH transcription factors that regulate jasmonate responses in Arabidopsis , 2011, Journal of experimental botany.

[26]  M. Berger,et al.  Universal protein-binding microarrays for the comprehensive characterization of the DNA-binding specificities of transcription factors , 2009, Nature Protocols.

[27]  V. Joardar,et al.  The Arabidopsis thaliana JASMONATE INSENSITIVE 1 gene is required for suppression of salicylic acid-dependent defenses during infection by Pseudomonas syringae. , 2006, Molecular plant-microbe interactions : MPMI.

[28]  Dirk Inzé,et al.  Boosting tandem affinity purification of plant protein complexes. , 2008, Trends in plant science.

[29]  D. Xie,et al.  COI1: an Arabidopsis gene required for jasmonate-regulated defense and fertility. , 1998, Science.

[30]  J. Franco-Zorrilla,et al.  Improved protein-binding microarrays for the identification of DNA-binding specificities of transcription factors. , 2011, The Plant journal : for cell and molecular biology.

[31]  A. Bauch,et al.  An efficient tandem affinity purification procedure for interaction proteomics in mammalian cells , 2006, Nature Methods.

[32]  M. Hamberg,et al.  (+)-7-iso-Jasmonoyl-L-isoleucine is the endogenous bioactive jasmonate. , 2009, Nature chemical biology.

[33]  B. Causier,et al.  The TOPLESS Interactome: A Framework for Gene Repression in Arabidopsis1[W][OA] , 2011, Plant Physiology.

[34]  J. Franco-Zorrilla,et al.  The Arabidopsis bHLH Transcription Factors MYC3 and MYC4 Are Targets of JAZ Repressors and Act Additively with MYC2 in the Activation of Jasmonate Responses[C][W] , 2011, Plant Cell.

[35]  Hoo Sun Chung,et al.  Negative Feedback Control of Jasmonate Signaling by an Alternative Splice Variant of JAZ101[C][W][OA] , 2013, Plant Physiology.

[36]  C. Wasternack,et al.  Hydroxylated jasmonates are commonly occurring metabolites of jasmonic acid and contribute to a partial switch-off in jasmonate signaling. , 2007, The New phytologist.

[37]  E. Esmans,et al.  Metabolic Fate of Jasmonates in Tobacco Bright Yellow-2 Cells1 , 2004, Plant Physiology.

[38]  R. Solano,et al.  The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. , 2009, The Plant journal : for cell and molecular biology.

[39]  T. Wada,et al.  Jasmonic acid control of GLABRA3 links inducible defense and trichome patterning in Arabidopsis , 2009, Development.

[40]  W. E. Hillis,et al.  The phenolic constituents of Prunus domestica. I.—The quantitative analysis of phenolic constituents , 1959 .

[41]  J. Rizo,et al.  Jasmonate perception by inositol phosphate-potentiated COI1-JAZ co-receptor , 2010, Nature.

[42]  Bryan C Thines,et al.  JAZ repressor proteins are targets of the SCFCOI1 complex during jasmonate signalling , 2007, Nature.

[43]  A. Goossens,et al.  The JAZ Proteins: A Crucial Interface in the Jasmonate Signaling Cascade , 2011, Plant Cell.

[44]  J. Turner,et al.  Wound-Induced Endogenous Jasmonates Stunt Plant Growth by Inhibiting Mitosis , 2008, PloS one.

[45]  A. J. Koo,et al.  A bHLH-Type Transcription Factor, ABA-INDUCIBLE BHLH-TYPE TRANSCRIPTION FACTOR/JA-ASSOCIATED MYC2-LIKE1, Acts as a Repressor to Negatively Regulate Jasmonate Signaling in Arabidopsis[C][W] , 2013, Plant Cell.

[46]  Zhiwei Cheng,et al.  The bHLH transcription factor MYC3 interacts with the Jasmonate ZIM-domain proteins to mediate jasmonate response in Arabidopsis. , 2011, Molecular plant.

[47]  G. Howe,et al.  New Weapons and a Rapid Response against Insect Attack1 , 2008, Plant Physiology.

[48]  G. Howe,et al.  Alternative splicing expands the repertoire of dominant JAZ repressors of jasmonate signaling. , 2010, The Plant journal : for cell and molecular biology.

[49]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[50]  D. Werck-Reichhart,et al.  Cytochromes P450 CYP94C1 and CYP94B3 Catalyze Two Successive Oxidation Steps of Plant Hormone Jasmonoyl-isoleucine for Catabolic Turnover , 2012, The Journal of Biological Chemistry.

[51]  Fang Wang,et al.  Jasmonate Regulates the INDUCER OF CBF EXPRESSION–C-REPEAT BINDING FACTOR/DRE BINDING FACTOR1 Cascade and Freezing Tolerance in Arabidopsis[W] , 2013, Plant Cell.

[52]  Hoo Sun Chung,et al.  A Critical Role for the TIFY Motif in Repression of Jasmonate Signaling by a Stabilized Splice Variant of the JASMONATE ZIM-Domain Protein JAZ10 in Arabidopsis[C][W] , 2009, The Plant Cell Online.

[53]  J. B. Reid,et al.  MYC2 Differentially Modulates Diverse Jasmonate-Dependent Functions in Arabidopsis[W] , 2007, The Plant Cell Online.

[54]  C. Wasternack,et al.  Jasmonates: biosynthesis, perception, signal transduction and action in plant stress response, growth and development. An update to the 2007 review in Annals of Botany. , 2013, Annals of botany.

[55]  Eric Bonnet,et al.  A Tandem Affinity Purification-based Technology Platform to Study the Cell Cycle Interactome in Arabidopsis thaliana*S , 2007, Molecular & Cellular Proteomics.

[56]  E. Weiler,et al.  Structure-activity analyses reveal the existence of two separate groups of active octadecanoids in elicitation of the tendril-coiling response of Bryonia dioica Jacq. , 1999, Planta.

[57]  R. Solano,et al.  Molecular players regulating the jasmonate signalling network. , 2005, Current opinion in plant biology.

[58]  D. Arnon COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. , 1949, Plant physiology.

[59]  W. Peng,et al.  The Jasmonate-ZIM Domain Proteins Interact with the R2R3-MYB Transcription Factors MYB21 and MYB24 to Affect Jasmonate-Regulated Stamen Development in Arabidopsis[C][W] , 2011, Plant Cell.

[60]  D. Gross,et al.  Pseudomonas syringae Phytotoxins: Mode of Action, Regulation, and Biosynthesis by Peptide and Polyketide Synthetases , 1999, Microbiology and Molecular Biology Reviews.

[61]  K. Mysore,et al.  The phytotoxin coronatine contributes to pathogen fitness and is required for suppression of salicylic acid accumulation in tomato inoculated with Pseudomonas syringae pv. tomato DC3000. , 2007, Molecular plant-microbe interactions : MPMI.

[62]  F. Harnisch,et al.  Independently silencing two JAR family members impairs levels of trypsin proteinase inhibitors but not nicotine , 2007, Planta.

[63]  H. Kawaide,et al.  Arabidopsis CYP94B3 encodes jasmonyl-L-isoleucine 12-hydroxylase, a key enzyme in the oxidative catabolism of jasmonate. , 2011, Plant & cell physiology.

[64]  I. Tiryaki,et al.  The Oxylipin Signal Jasmonic Acid Is Activated by an Enzyme That Conjugates It to Isoleucine in Arabidopsis , 2004, The Plant Cell Online.