Abstract
Despite the importance of the senescence processes in plants, our knowledge on regulatory mechanisms of senescence is still poor. WRKY transcription factors have been shown to be involved in the regulation of leaf senescence. However, almost nothing is known about the upstream regulation of the senescence specific expression of WRKY factors. Therefore, we characterized proteins that bind and activate the promoter of WRKY53, which participates in leaf senescence in Arabidopsis thaliana. Surprisingly, a mitogen activated protein kinase kinase kinase (MEKK1) was identified as a DNA-binding protein. The binding motif for MEKK1 in the WRKY53 promoter could be characterized and promoter:GUS analyses revealed that this region is important for the switch of WRKY53 expression from a leaf age dependent to a systemic plant age dependent expression during bolting time. In addition to its promotor-binding activity, MEKK1 was also able to interact with the WRKY53 protein. Using bimolecular fluorescence complementation assays the complex formation of MEKK1 and WRKY53 could be localized predominately in the nucleus of Arabidopsis cells. MEKK1 could also phosphorylate WRKY53 in vitro and phosphorylation could increase DNA-binding activity of WRKY53 in vitro and transcription of a WRKY53 promoter driven reporter gene in vivo. These results suggest that MEKK1 is a bifunctional protein: it binds to the promoter of the WRKY53 gene regulating the switch from a leaf age dependent to a plant age dependent expression and it can phosopharylate WRKY53 in vitro increasing its DNA binding activity. Thus, MEKK1 might be able to take a very direct short cut in mitogen-activated protein kinase (MAPK) signalling by directly phosphorylating a transcription factor.
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Andreasson E, Jenkins T, Brodersen P, Thorgrimsen S, Petersen NHT, Zhu SJ, Qiu JL, Micheelsen P, Rocher A, Petersen M, Newman MA, Nielsen HB, Hirt H, Somssich I, Mattsson O, Mundy J (2005) The MAP kinase substrate MKS1 is a regulator of plant defense responses. EMBO J 24:2579–2589
Arrigo AP (1999) Gene expression and the thiol redox state. Free Radical Biol Med 27:936–944
Asai T, Tena G, Plotnikova J, Willmann MR, Chiu W-L, Gomez-Gomez L, Boller T, Ausubel FM, Sheen J (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983
Batoko H, Zheng HQ, Hawes C, Moore I (2000) A rab1 GTPase is required for transport between the endoplasmic reticulum and golgi apparatus and for normal golgi movement in plants. Plant Cell 12:2201–2218
Bechtold N, Pelletier G (1998) In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Methods Mol Biol 82:259–266
Buchanan-Wollaston V, Earl S, Harrison E, Mathas E, Navabpour S, Page T, Pink D (2003) The molecular analysis of leaf senescence—a genomics approach. Plant Biotech J 1:3–22
Buchanan-Wollaston V, Page T, Harrison E, Breeze E, Lim PO, Nam HG, Lin JF, Wu SH, Swidzinski J, Ishizaki K, Leaver CJ (2005) Comparative transcriptome analysis reveals significant differences in gene expression and signalling pathways between developmental and dark/starvation-induced senescence in Arabidopsis. Plant J 42:567–585
Cheong YH, Moon BC, Kim JK, Kim CY, Kim MC, Kim IH, Park CY, Kim JC, Park BO, Koo SC, Yoon HW, Chung WS, Lim CO, Lee SY, Cho MJ (2003) BWMK1, a rice mitogen-activated protein kinase, locates in the nucleus and mediates pathogenesis-related gene expression by activation of a transcription factor. Plant Physiol 132:1961–1972
Choi G, Kim J-I, Hong S-W, Shin B, Choi G, Blakeslee JJ, Murphy AS, Seo YW, Kim K, Koh E-J, Song P-S, Lee H (2005) A possible role for NDPK2 in the regulation of auxin-mediated responses for plantgrowth and development. Plant Cell Physiol 46:1246–1254
Dong J, Chen C, Chen Z (2003) Expression profile of the WRKY gene superfamily during plant defence. Plant Mol Biol 51:21–37
Edmunds JW, Mahadevan LC (2004) MAP kinases as structural adaptors and enzymatic activators in transcription complexes. J Cell Sci 117:3715–3723
Edmunds JW, Mahadevan LC (2006) Cell signaling. Protein kinases seek close encounters with active genes. Science 313:449–451
Eulgem T, Rushton PJ, Robatzek S, Somssich IE (2000) The WRKY superfamily of plant transcription factors. Trends Plant Sci 5:199–206
Guo Y, Cai Z, Gan S (2004) Transcriptome of Arabidopsis leaf senescence. Plant Cell Environ 27:521–549
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory elements (PLACE) database. Nucl Acid Res 27:297–300
Hinderhofer K, Zentgraf U (2001) Identification of a transcription factor specifically expressed at the onset of leaf senescence. Planta 213:469–473
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Lee J, Rudd JJ, Macioszek VK, Scheel D (2004) Dynamic changes in the localization of MAPK cascade components controlling pathogenesis-related (PR) gene expression during innate immunity in parsley. J Biol Chem 279:22440–22448
Li J, Brader G, Palva ET (2004) The WRKY70 transcription factor: a node of convergence of jasmonate-mediated and salicylate-mediated signals in plant defense. Plant Cell 16:319–331
Lin JF, Wu SH (2004) Molecular events in senescing Arabidopsis leaves. Plant J 39:612–628
MAPK Group (Kazuya Ichimura et al) (2002) Mitogen-activated-protein-kinase cascades in plants: new nomenclature. Trends Plant Sci 7:301–308
Miao Y, Laun T, Zimmermann P, Zentgraf U (2004) Targets of the WRKY53 transcription factor and its role during leaf senescence in Arabidopsis. Plant Mol Biol 55:853–867
Miller JD, Arteca RN, Pell EJ (1999) Senescence-associated gene expression during ozone-induced leaf senescence in Arabidopsis. Plant Physiol 120:1015–1023
Moon H, Lee B, Choi G, Shin D, Prasad DT, Lee O, Kwak SS, Kim DH, Nam J, Bahk J, Hong JC, Lee SY, Cho MJ, Lim CO, Yun DJ (2003) NDP kinase 2 interacts with two oxidative stress-activated MAPKs to regulate cellular redox state and enhances multiple stress tolerance in transgenic plants. Proc Natl Acad Sci USA 100:358–363
Morrison DK, Davis RJ (2003) Regulation of MAP kinase signaling modules by scaffold proteins in mammals. Annu Rev Cell Dev Biol 19:91–118
Nakagami H, Kiegerl S, Hirt H (2004) OMTK1, a novel MAPKKK, channels oxidative stress signalling through direct MAPK interaction. J Biol Chem 27:26959–26966
Nakagami H, Pitschke A, Hirt H (2005) Emerging MAP kinase pathways in plant stress signalling. Trends Plant Sci 10:339–346
Nakagami H, Soukupova H, Schikora A, Zarsky V, Hirt H (2006) A mitogen-activated protein kinase kinase kinase mediates reactive oxygen species homeostasis in Arabidopsis. J Biol Chem 281:38697–38704
Navabpour S, Morris A, Allen R, Harrison E, Mackerness SAH, Buchanan-Wollaston V (2003) Expression of senescence–enhanced genes in response to oxidative stress. J Exp Bot 54:2285–2292
Pokholok DK, Zeitlinge J, Hannett NM, Reynolds DB, Young RA (2006) Activated signal transduction kinases frequently occupy target genes. Science 313:533–536
Robatzek S, Somssich IE (2002) Targets of AtWRKY6 regulation during plant senescence and pathogen defense. Genes Dev 16:1139–1149
Sheen J (2001) Signal transduction in Maize and Arabidopsis mesophyll protoplasts. Plant Physiol 127:1466–1475
Tang DZ, Innes RW (2002) Overexpression of a kinase-deficient form of the EDR1 gene enhances powdery mildew resistance and ethylene-induced senescence in Arabidopsis. Plant J 32:975–983
Teige M, Scheikl E, Eulgem T, Doczi F, Ichimura K, Shinozaki K, Dangl JL, Hirt H (2004) The MKK2 pathway mediates cold and salt stress signaling in Arabidopsis. Mol Cell 15:141–152
Ülker B, Somssich IE (2004) WRKY factors: from DNA binding towards biological function. Curr Opin Plant Biol 7:491–498
Wan JR, Zhang SQ, Stacey G (2004) Activation of a mitogen-activated protein kinase pathway in Arabidopsis by chitin. Mol Plant Path 5:125–135
Wrzaczek M, Hirt H (2001) Plant MAP kinase pathways: how many and what for? Biol Cell 93:81–87
Xiang CB, Han P, Lutziger I, Wang K, Oliver DJ (1999) A mini binary vector series for plant transformation. Plant Mol Biol 40:711–717
Ye ZZ, Rodriguez R, Tran A, Hoang H, de los Santos D, Brown S, Vellanoweth RL (2000) The developmental transition to flowering represses ascorbate peroxidase activity and induces enzymatic lipid peroxidation in leaf tissue in Arabidopsis thaliana. Plant Sci 158:115–127
Zentgraf U, Kolb D, Laun T, Rentsch D (2004) Senescence related gene expression profiles of rosette leaves of Arabidopsis thaliana: leaf age versus plant age. Plant Biol 6:178–183
Zimmermann P, Orendi G, Heinlein C, Zentgraf U (2006) Senescence specific regulation of catalases in Arabidopsis thaliana (L.) Heynh. Plant Cell Environ 29:1049–1060
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We acknowledge the Nottingham Arabidopsis Stock Centre (NASC) for providing the seeds of the T-DNA insertion lines and the DFG (SFB446) for financial support.
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Miao, Y., Laun, T.M., Smykowski, A. et al. Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter. Plant Mol Biol 65, 63–76 (2007). https://doi.org/10.1007/s11103-007-9198-z
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DOI: https://doi.org/10.1007/s11103-007-9198-z