The CRY2-COP1-HY5-BBX7/8 module regulates blue light-dependent cold acclimation in Arabidopsis.

Light and temperature are two key environmental factors that coordinately regulate plant growth and development. Although the mechanisms that integrate signaling mediated by cold and red light have been unraveled, the roles of the blue light photoreceptors cryptochromes in plant responses to cold remain unclear. In this study, we demonstrate that the CRYPTOCHROME2 (CRY2)-COP1-HY5-BBX7/8 module regulates blue light-dependent cold acclimation in Arabidopsis thaliana. We show that phosphorylated forms of CRY2 induced by blue light are stabilized by cold stress and that cold-stabilized CRY2 competes with the transcription factor HY5 to attenuate the HY5-COP1 interaction, thereby allowing HY5 to accumulate at cold temperatures. Furthermore, our data demonstrate that B-BOX DOMAIN PROTEIN7 (BBX7) and BBX8 function as direct HY5 targets that positively regulate freezing tolerance by modulating the expression of a set of cold-responsive genes, which mainly occurs independently of the C-REPEAT-BINDING FACTOR pathway. Our study uncovers a mechanistic framework by which CRY2-mediated blue-light signaling enhances freezing tolerance, shedding light on the molecular mechanisms underlying the crosstalk between cold and light signaling pathways in plants.

[1]  Z. Gong,et al.  The direct targets of CBFs: in cold stress response and beyond. , 2021, Journal of integrative plant biology.

[2]  Margaret Ahmad,et al.  Effect of temperature on the Arabidopsis cryptochrome photocycle. , 2021, Physiologia plantarum.

[3]  J. Wohlschlegel,et al.  Regulation of Arabidopsis photoreceptor CRY2 by two distinct E3 ubiquitin ligases , 2020, Nature Communications.

[4]  T. Schmülling,et al.  Light acts as a stressor and influences abiotic and biotic stress responses in plants. , 2020, Plant, cell & environment.

[5]  Miriam Lohr,et al.  Identification of BBX proteins as rate-limiting cofactors of HY5 , 2020, Nature Plants.

[6]  Jigang Li,et al.  Cold-Induced CBF-PIF3 Interaction Enhances Freezing Tolerance by Stabilizing the phyB Thermosensor in Arabidopsis. , 2020, Molecular plant.

[7]  Dongqing Xu,et al.  B-box proteins: pivotal players in light-mediated development in plants. , 2020, Journal of integrative plant biology.

[8]  W. Terzaghi,et al.  Modulation of BIN2 kinase activity by HY5 controls hypocotyl elongation in the light , 2020, Nature Communications.

[9]  Chentao Lin,et al.  Mechanisms of Cryptochrome-Mediated Photoresponses in Plants. , 2020, Annual review of plant biology.

[10]  T. Ritz,et al.  Cryptochrome mediated magnetic sensitivity in Arabidopsis occurs independently of light-induced electron transfer to the flavin , 2020, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[11]  Yiting Shi,et al.  Molecular Regulation of Plant Responses to Environmental Temperatures. , 2020, Molecular plant.

[12]  U. Hoecker,et al.  Cryptochrome 2 competes with COP1 substrates to repress COP1 ubiquitin ligase activity during Arabidopsis photomorphogenesis , 2019, Proceedings of the National Academy of Sciences.

[13]  Dongqing Xu COP1- and BBXs-HY5-Mediated Light Signal Transduction in Plants. , 2019, The New phytologist.

[14]  C. Fankhauser,et al.  Molecular mechanisms underlying phytochrome-controlled morphogenesis in plants , 2019, Nature Communications.

[15]  Steven L Salzberg,et al.  Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype , 2019, Nature Biotechnology.

[16]  X. Deng,et al.  B-Box Containing Proteins BBX30 and BBX31, Acting Downstream of HY5, Negatively Regulate Photomorphogenesis in Arabidopsis1 , 2019, Plant Physiology.

[17]  Maneesh Lingwan,et al.  The B-Box-Containing MicroProtein miP1a/BBX31 Regulates Photomorphogenesis and UV-B Protection1[OPEN] , 2019, Plant Physiology.

[18]  Meng Chen,et al.  Daytime temperature is sensed by phytochrome B in Arabidopsis through a transcriptional activator HEMERA , 2019, Nature Communications.

[19]  X. Deng,et al.  B-BOX DOMAIN PROTEIN28 Negatively Regulates Photomorphogenesis by Repressing the Activity of Transcription Factor HY5 and Undergoes COP1-Mediated Degradation , 2018, Plant Cell.

[20]  Yiting Shi,et al.  Molecular Regulation of CBF Signaling in Cold Acclimation. , 2018, Trends in plant science.

[21]  S. Datta,et al.  Two B-Box Proteins Regulate Photomorphogenesis by Oppositely Modulating HY5 through their Diverse C-Terminal Domains1[OPEN] , 2018, Plant Physiology.

[22]  S. Hasegawa,et al.  Phototropin perceives temperature based on the lifetime of its photoactivated state , 2017, Proceedings of the National Academy of Sciences.

[23]  Jigang Li,et al.  PIF3 is a negative regulator of the CBF pathway and freezing tolerance in Arabidopsis , 2017, Proceedings of the National Academy of Sciences.

[24]  Rongcheng Lin,et al.  A PIF1/PIF3-HY5-BBX23 Transcription Factor Cascade Affects Photomorphogenesis1[OPEN] , 2017, Plant Physiology.

[25]  Yiting Shi,et al.  BZR1 Positively Regulates Freezing Tolerance via CBF-Dependent and CBF-Independent Pathways in Arabidopsis. , 2017, Molecular plant.

[26]  James C. W. Locke,et al.  Phytochromes function as thermosensors in Arabidopsis , 2016, Science.

[27]  E. Schäfer,et al.  Phytochrome B integrates light and temperature signals in Arabidopsis , 2016, Science.

[28]  Wei Liu,et al.  Photoactivation and inactivation of Arabidopsis cryptochrome 2 , 2016, Science.

[29]  Z. Gong,et al.  The cbfs triple mutants reveal the essential functions of CBFs in cold acclimation and allow the definition of CBF regulons in Arabidopsis. , 2016, The New phytologist.

[30]  B. Liu,et al.  The Blue Light-Dependent Polyubiquitination and Degradation of Arabidopsis Cryptochrome2 Requires Multiple E3 Ubiquitin Ligases. , 2016, Plant & cell physiology.

[31]  X. Deng,et al.  BBX21, an Arabidopsis B-box protein, directly activates HY5 and is targeted by COP1 for 26S proteasome-mediated degradation , 2016, Proceedings of the National Academy of Sciences.

[32]  Jian-Kang Zhu,et al.  Mutational Evidence for the Critical Role of CBF Transcription Factors in Cold Acclimation in Arabidopsis1 , 2016, Plant Physiology.

[33]  J. Ecker,et al.  Cryptochromes Interact Directly with PIFs to Control Plant Growth in Limiting Blue Light , 2016, Cell.

[34]  J. Noel,et al.  Cryptochrome 1 interacts with PIF4 to regulate high temperature-mediated hypocotyl elongation in response to blue light , 2015, Proceedings of the National Academy of Sciences.

[35]  Colleen J Doherty,et al.  Regulation of the Arabidopsis CBF regulon by a complex low-temperature regulatory network. , 2015, The Plant journal : for cell and molecular biology.

[36]  Qian Luo,et al.  Red-light-dependent interaction of phyB with SPA1 promotes COP1-SPA1 dissociation and photomorphogenic development in Arabidopsis. , 2015, Molecular plant.

[37]  Q. Xie,et al.  OST1 kinase modulates freezing tolerance by enhancing ICE1 stability in Arabidopsis. , 2015, Developmental cell.

[38]  Ling Zhu,et al.  Light-Activated Phytochrome A and B Interact with Members of the SPA Family to Promote Photomorphogenesis in Arabidopsis by Reorganizing the COP1/SPA Complex , 2015, Plant Cell.

[39]  E. Schäfer,et al.  Molecular mechanisms for mediating light-dependent nucleo/cytoplasmic partitioning of phytochrome photoreceptors , 2014, The New phytologist.

[40]  T. Kottke,et al.  Proton Transfer to Flavin Stabilizes the Signaling State of the Blue Light Receptor Plant Cryptochrome* , 2014, The Journal of Biological Chemistry.

[41]  Xuecheng Wang,et al.  A CRISPR/Cas9 toolkit for multiplex genome editing in plants , 2014, BMC Plant Biology.

[42]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[43]  J. Botto,et al.  The BBX family of plant transcription factors. , 2014, Trends in plant science.

[44]  Anton J. Enright,et al.  Kraken: A set of tools for quality control and analysis of high-throughput sequence data , 2013, Methods.

[45]  Wei Shi,et al.  featureCounts: an efficient general purpose program for assigning sequence reads to genomic features , 2013, Bioinform..

[46]  R. Bittl,et al.  Lifetimes of Arabidopsis cryptochrome signaling states in vivo. , 2013, The Plant journal : for cell and molecular biology.

[47]  J. Botto,et al.  The Arabidopsis B-BOX Protein BBX25 Interacts with HY5, Negatively Regulating BBX22 Expression to Suppress Seedling Photomorphogenesis[C][W] , 2013, Plant Cell.

[48]  Jeongmoo Park,et al.  Phytochrome B inhibits binding of phytochrome-interacting factors to their target promoters. , 2012, The Plant journal : for cell and molecular biology.

[49]  M. Thomashow,et al.  Photoperiodic regulation of the C-repeat binding factor (CBF) cold acclimation pathway and freezing tolerance in Arabidopsis thaliana , 2012, Proceedings of the National Academy of Sciences.

[50]  R. Catalá,et al.  Integration of low temperature and light signaling during cold acclimation response in Arabidopsis , 2011, Proceedings of the National Academy of Sciences.

[51]  Xuanming Liu,et al.  Blue Light-Dependent Interaction of CRY2 with SPA1 Regulates COP1 activity and Floral Initiation in Arabidopsis , 2011, Current Biology.

[52]  B. Liu,et al.  Arabidopsis cryptochrome 1 interacts with SPA1 to suppress COP1 activity in response to blue light. , 2011, Genes & development.

[53]  Yan-chun Zhang,et al.  Blue-light-dependent interaction of cryptochrome 1 with SPA1 defines a dynamic signaling mechanism. , 2011, Genes & development.

[54]  Haiyang Wang,et al.  Arabidopsis Transcription Factor ELONGATED HYPOCOTYL5 Plays a Role in the Feedback Regulation of Phytochrome A Signaling[C][W] , 2010, Plant Cell.

[55]  Jie Huang,et al.  Formation of Nuclear Bodies of Arabidopsis CRY2 in Response to Blue Light Is Associated with Its Blue Light–Dependent Degradation[W] , 2009, The Plant Cell Online.

[56]  V. Rubio,et al.  LZF1/SALT TOLERANCE HOMOLOG3, an Arabidopsis B-Box Protein Involved in Light-Dependent Development and Gene Expression, Undergoes COP1-Mediated Ubiquitination[W] , 2008, The Plant Cell Online.

[57]  J. Kudla,et al.  In Planta Visualization of Protein Interactions Using Bimolecular Fluorescence Complementation (BiFC). , 2008, CSH protocols.

[58]  Yujing Wang,et al.  Firefly Luciferase Complementation Imaging Assay for Protein-Protein Interactions in Plants1[C][W][OA] , 2007, Plant Physiology.

[59]  G. Whitelam,et al.  Light-quality regulation of freezing tolerance in Arabidopsis thaliana , 2007, Nature Genetics.

[60]  K. Miura,et al.  SIZ1-Mediated Sumoylation of ICE1 Controls CBF3/DREB1A Expression and Freezing Tolerance in Arabidopsis[W][OA] , 2007, The Plant Cell Online.

[61]  P. Quail,et al.  Photoactivated phytochrome induces rapid PIF3 phosphorylation prior to proteasome-mediated degradation. , 2006, Molecular cell.

[62]  R. Hellens,et al.  Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants , 2005, Plant Methods.

[63]  Y. Sang,et al.  From The Cover: A role for Arabidopsis cryptochromes and COP1 in the regulation of stomatal opening. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[64]  V. Rubio,et al.  The COP1-SPA1 interaction defines a critical step in phytochrome A-mediated regulation of HY5 activity. , 2003, Genes & development.

[65]  D. Weigel,et al.  A thermosensory pathway controlling flowering time in Arabidopsis thaliana , 2003, Nature Genetics.

[66]  T. Mockler,et al.  Regulation of Arabidopsis cryptochrome 2 by blue-light-dependent phosphorylation , 2002, Nature.

[67]  B. Winkel-Shirley,et al.  Biosynthesis of flavonoids and effects of stress. , 2002, Current opinion in plant biology.

[68]  Michael F. Thomashow,et al.  PLANT COLD ACCLIMATION: Freezing Tolerance Genes and Regulatory Mechanisms. , 1999, Annual review of plant physiology and plant molecular biology.

[69]  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.

[70]  P. Quail,et al.  PIF3, a Phytochrome-Interacting Factor Necessary for Normal Photoinduced Signal Transduction, Is a Novel Basic Helix-Loop-Helix Protein , 1998, Cell.

[71]  X. Deng,et al.  Multiple photoreceptors mediate the light-induced reduction of GUS-COP1 from Arabidopsis hypocotyl nuclei. , 1998, The Plant journal : for cell and molecular biology.

[72]  K. Shinozaki,et al.  Two Transcription Factors, DREB1 and DREB2, with an EREBP/AP2 DNA Binding Domain Separate Two Cellular Signal Transduction Pathways in Drought- and Low-Temperature-Responsive Gene Expression, Respectively, in Arabidopsis , 1998, Plant Cell.

[73]  E. Stockinger,et al.  Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[74]  S. Gelvin,et al.  Strength and tissue specificity of chimeric promoters derived from the octopine and mannopine synthase genes , 1995 .

[75]  X. Deng,et al.  Light inactivation of arabidopsis photomorphogenic repressor COP1 involves a cell-specific regulation of its nucleocytoplasmic partitioning , 1994, Cell.

[76]  Bing-kai Hou,et al.  The Arabidopsis UDP‐glycosyltransferases UGT79B2 and UGT79B3, contribute to cold, salt and drought stress tolerance via modulating anthocyanin accumulation , 2017, The Plant journal : for cell and molecular biology.

[77]  Haiyang Wang,et al.  Phytochrome Signaling Mechanisms , 2011, The arabidopsis book.

[78]  P. Quail,et al.  PIFs: pivotal components in a cellular signaling hub. , 2011, Trends in plant science.

[79]  L. Lepiniec,et al.  Flavonoid oxidation in plants: from biochemical properties to physiological functions. , 2007, Trends in plant science.

[80]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .