Abstract
Key message
Arabidopsis BIK1 negatively regulates EIN3-depedent gene expression as an immediate cellular response. BIK1 localizes to the plasma membrane and its autophosphorylation and kinase activity involves in EIN3 repression.
Abstract
BOTRYTIS INDUCED KINASE1 (BIK1) is a multifunctional receptor-like kinase that involves in ethylene-mediated plant defense signaling. The loss of function BIK1 becomes insensitive to ethylene, but it still accumulates a higher level of ETHYLENE INSENSITIVE3 (EIN3) that serves as the key transcription activator in ethylene signaling. To unequivocally elucidate BIK1 function on EIN3 regulation in ethylene signaling, we took a combined approach of transient expression assay and stable expression analysis of BIK1. In our cell-based functional assay BIK1 destabilized EIN3 and down-regulated EIN3-dependent transcription. Membrane localization and autophosphorylation of BIK1 were required for full repression of EIN3 function, but its kinase activity potential compromised such regulatory action. Consistently, the analysis of transgenic plants verified BIK1 function on EIN3 repression. Our findings have clarified that autophosphorylated BIK1 in the plasma membrane negatively regulates EIN3-dependent gene expression. Thus, ethylene insensitivity in bik1 appears to be an indirect or a feedback long-term response.
Similar content being viewed by others
Abbreviations
- BAK1:
-
BRI1-ASSOCIATED KINASE1
- BIK1:
-
BOTRYTIS INDUCED KINASE1
- BRI1:
-
BRASSINOLIDE INSENSITIVE1
- DEX:
-
Dexamethasone
- EIN3:
-
ETHYLENE INSENSITIVE3
- GR:
-
Glucocorticoid receptor
- RT-PCR:
-
Reverse transcriptase-dependent polymerase chain reaction
- SDS:
-
Sodium dodecyl sulfate
References
Abu Qamar S, Chai M, Luo H, Song F, Mengiste T (2008) Tomato protein kinase 1b mediates signaling of plant responses to necrotrophic fungi and insect herbivory. Plant Cell 20:1964–1983
Binder BM, Walker JM, Gagne JM, Emborg TJ, Hemmann G, Bleecker AB, Vierstra RD (2007) The Arabidopsis EIN3 binding F-Box proteins EBF1 and EBF2 have distinct but overlapping roles in ethylene signaling. Plant Cell 19:509–523
Boutin JA (1997) Myristoylation. Cell Signal 9:15–35
Chao Q, Rothenberg M, Solano R, Roman G, Terzaghi W, Ecker JR (1997) Activation of the ethylene gas response pathway in Arabidopsis by the nuclear protein Ethylene-insensitive 3 and related proteins. Cell 89:1133–1144
Cho YH, Yoo SD (2015) Novel connections and gaps in ethylene signaling from the ER membrane to the nucleus. Front Plant Sci 5:733. doi:10.3389/fpls.2014.00733
Cho YH, Hong JW, Kim EC, Yoo SD (2012) Regulatory functions of SnRK1 in stress-responsive gene expression and in plant growth and development. Plant Physiol 158:1955–1964
Gagne JM, Smalle J, Gingerich DJ, Walker JM, Yoo SD, Yanagisawa S, Vierstra RD (2004) Arabidopsis EIN3-binding F-box 1 and 2 form ubiquitin-protein ligases that repress ethylene action and promote growth by directing EIN3 degradation. Proc Natl Acad Sci USA 101:6803–6808
Guo H, Ecker JR (2003) Plant responses to ethylene gas are mediated by SCFEBF1/EBF2-dependent proteolysis of EIN3 transcription factor. Cell 115:667–677
Huffaker A, Ryan CA (2007) Endogenous peptide defense signals in Arabidopsis differentially amplify signaling for the innate immune response. Proc Natl Acad Sci USA 104:10732–10736
Im JH, Cho YH, Hong JW, Kang GH, Kim GD, Yoo SD (2014) Inverse modulation of the energy sensor Snf1-Related Protein Kinase1 on hypoxia adaptation and salt stress tolerance in Arabidopsis thaliana. Plant Cell Environ 37:2303–2312
Ju C, Yoon G, Shemansky J, Lin D, Ying ZI, Chang J, Garrett W, Kessenbrock M, Groth G, Tucker ML, Cooper B, Kieber JJ, Chang C (2012) CTR1 phosphorylates the central regulator EIN2 to control ethylene hormone signaling from the ER membrane to the nucleus in Arabidopsis. Proc Natl Acad Sci USA 109:19486–19491
Kim GD, Cho YH, Yoo SD (2015) Regulatory functions of evolutionarily conserved AN1/A20-like Zinc finger family proteins in Arabidopsis stress responses under high temperature. Biochem Biophys Res Commun 457:213–220
Laluk K, Luo H, Chai M, Dhawan R, Lai Z, Mengiste T (2011) Biochemical and genetic requirements for function of the immune response regulator BOTRYTIS-INDUCED KINASE1 in plant growth, ethylene signaling, and pamp triggered immunity in Arabidopsis. Plant Cell 23:2831–2849
Li JF, Chung HS, Niu Y, Bush J, McCormack M, Sheen J (2013) Comprehensive protein-based artificial microRNA screens for effective gene silencing in plants. Plant Cell 25:1507–1522
Lin W, Li B, Lu D, Chen S, Zhu N, He P, Shan L (2012) Tyrosine phosphorylation of protein kinase complex BAK1/BIK1 mediates Arabidopsis innate immunity. Proc Natl Acad Sci USA 111:3632–3637
Lin W, Lu D, Gao X, Jiang S, Ma X, Wang Z, Mengiste T, He P, Shan L (2013) Inverse modulation of plant immune and brassinosteroid signaling pathways by the receptor-like cytoplasmic kinase BIK1. Proc Natl Acad Sci USA 110:12114–12119
Liu Z, Wu Y, Yang F, Zhang Y, Chen S, Xie Q, Tian X, Zhou JM (2012) BIK1 interacts with PEPRs to mediate ethylene-induced immunity. Proc Natl Acad Sci USA 110:6205–6210
Lua D, Wub S, Gaoa X, Zhangb Y, Shanb L, He P (2009) A receptor-like cytoplasmic kinase, BIK1, associates with a flagellin receptor complex to initiate plant innate immunity. Proc Natl Acad Sci USA 107:496–501
Potuschak T, Lechner E, Parmentier Y, Yanagisawa S, Grava S, Koncz C, Genschik P (2003) EIN3-dependent regulation of plant ethylene hormone signaling by two Arabidopsis F box proteins: EBF1 and EBF2. Cell 115:679–689
Qiao H, Shen Z, Huang SSC, Schmitz RJ, Urich MA, Briggs SP, Ecker JR (2012) Processing and subcellular trafficking of ER-tethered EIN2 control response to ethylene gas. Science 338:390–393
Taylor SS, Zhang P, Steichen JM, Keshwani MM, Kornev AP (2013) PKA: lessons learned after twenty years. Biochim Biophys Acta 1834:1271–1278
Veronese P, Nakagami H, Bluhm B, AbuQamar S, Chen X, Salmeron J, Dietrich RA, Hirt H, Mengistea T (2006) The membrane-anchored BOTRYTIS-INDUCED KINASE1 plays distinct roles in Arabidopsis resistance to necrotrophic and biotrophic pathogens. Plant Cell 18:257–273
Wen X, Zhang C, Ji Y, Zhao Q, He W, An F, Jiang L, Guo H (2012) Activation of ethylene signaling is mediated by nuclear translocation of the cleaved EIN2 carboxyl terminus. Cell Res 22:1613–1616
Yanagisawa S, Yoo SD, Sheen J (2003) Differential regulation of EIN3 stability by glucose and ethylene signalling in plants. Nature 425:521–525
Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572
Acknowledgments
This study was supported by the Korean NRF (NRF-2012R1A2A2A01005933) and the Woo Jang Chun Special Project funded by the Korean RDA (PJ009106052014) to SDY.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by J. Register.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Kang, GH., Son, S., Cho, YH. et al. Regulatory role of BOTRYTIS INDUCED KINASE1 in ETHYLENE INSENSITIVE3-dependent gene expression in Arabidopsis . Plant Cell Rep 34, 1605–1614 (2015). https://doi.org/10.1007/s00299-015-1812-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-015-1812-y