A HECT E3 ubiquitin ligase negatively regulates Arabidopsis leaf senescence through degradation of the transcription factor WRKY53.
WRKY transcription factors play a central role in controlling leaf senescence in Arabidopsis. One important member, WRKY53, is tightly regulated by various mechanisms, and is a convergence node between senescence and pathogen responses. Using WRKY53 in a yeast two-hybrid screen, we isolated the HECT domain E3 ubiquitin ligase UPL5. In contrast to mammals, Arabidopsis contains only seven HECT E3 ubiquitin ligases, whose targets and functions are largely unknown. In yeast cells, UPL5 interacts with WRKY53 via its leucine zipper domain, and this interaction was confirmed in the cytoplasm of plant cells by a bimolecular fluorescence complementation assay. UPL5 was able to use the WRKY53 protein as a substrate for polyubiquitination in an in vitro system, and induction of UPL5 expression by an ethanol-inducible system in upl5 plants led to degradation of the WRKY53 protein. Expression of both genes is regulated antagonistically in response to hydrogen peroxide, jasmonic acid and plant development. Two T-DNA insertion lines (upl5-1 and upl5-2) showed the same senescence phenotype as WRKY53 over-expressers. Over-expression of WRKY53 in the upl5 background enhanced the accelerated senescence phenotype of WRKY53 over-expressers. Therefore, we conclude that UPL5 regulates leaf senescence in Arabidopsis through degradation of WRKY53 and ensures that senescence is executed in the correct time frame.
Miao
Ying
Y
Zentgraf
Ulrike
U
D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D016254:Mutagenesis, Insertional; D018515:Plant Leaves / Q000254:growth & development*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D018749:RNA, Plant / Q000235:genetics; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination
D029681:Arabidopsis Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D018749:RNA, Plant; C036483:T-DNA; C496136:WRKY53 protein, Arabidopsis; D044767:Ubiquitin-Protein Ligases
Ay, N., Irmler, K., Fischer, A., Uhlemann, R., Reuter, G. and Humbeck, K. (2009) Epigenetic programming via histone methylation at WRKY53 controls leaf senescence in Arabidopsis thaliana. Plant J. 58, 333-346.
Balazadeh, S., Riaño-Pachón, D.M. and Mueller-Roeber, B. (2008) Transcription factors regulating leaf senescence in Arabidopsis thaliana. Plant Biol. 10 (Suppl. 1), 63-75.
Bechtold, N. and 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. and 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. et al. (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.
Chen, W.Q., Provart, N.J., Glazebrook, J. et al. (2002) Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. Plant Cell, 14, 559-574.
Dong, J., Chen, C. and Chen, Z. (2003) Expression profile of the WRKY gene superfamily during plant defence. Plant Mol. Biol. 51, 21-37.
Downes, B.P., Stupar, R.M., Gingerich, D.J. and Vierstra, R.D. (2003) The HECT ubiquitin-protein ligase (UPL) family in Arabidopsis: UPL3 has a specific role in trichome development. Plant J. 35, 729-742.
El-Refy, A., Perazza, D., Zekraoui, L., Valay, J.G., Bechtold, N., Brown, S., Hülskamp, M., Herzog, M. and Bonneville, J.M. (2003) The Arabidopsis KAKTUS gene encodes a HECT protein and controls the number of endoreduplication cycles. Mol. Genet. Genomics, 270, 403-414.
Eulgem, T., Rushton, P.J., Robatzek, S. and Somssich, I.E. (2000) The WRKY superfamily of plant transcription factors. Trends Plant Sci. 5, 199-206.
Gagne, J.M., Downes, B.P., Shiu, S.H., Dursk, A.M. and Vierstra, R.D. (2002) The F-box subunit of the SCF E3 complex is encoded by a diverse superfamily of genes in Arabidopsis. Proc. Natl Acad. Sci. USA, 99, 11519-11524.
Guo, Y., Cai, Z. and Gan, S. (2004) Transcriptome of Arabidopsis leaf senescence. Plant Cell Environ. 27, 521-549.
He, X.J., Hsu, Y.F., Zhu, S., Liu, H.L., Pontes, O., Zhu, J., Cui, X., Wang, C.S. and Zhu, J.K. (2009) A conserved transcriptional regulator is required for RNA-directed DNA methylation and plant development. Genes Dev. 23, 2717-2722.
Heazlewood, J.L., Tonti-Filippini, J.S., Gout, A.M., Day, D.A., Whelan, J. and Millar, A.H. (2004) Experimental analysis of the Arabidopsis mitochondrial proteome highlights signaling and regulatory components, provides assessment of targeting prediction programs, and indicates plant-specific mitochondrial proteins. Plant Cell, 16, 241-256.
Hinderhofer, K. and Zentgraf, U. (2001) Identification of a transcription factor specifically expressed at the onset of leaf senescence. Planta, 213, 469-473.
Huibregtse, J.M., Scheffner, M., Beaudenon, S. and Howley, P.M. (1995) A family of proteins structurally and functionally related to the E6-AP ubiquitin-protein ligase. Proc. Natl Acad. Sci. USA, 92, 2563-2567.
Jiang, Y. and Deyholos, M.K. (2009) Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses. Plant Mol. Biol. 69, 91-105.
Kosarev, P., Mayer, K.F.X. and Hardtke, C.S. (2002) Evaluation and classification of RING-finger domains encoded by the Arabidopsis genome. Genome Biol. 3, 1-12.
Krause, K., Kilbienski, I., Mulisch, M., Rödiger, A., Schäfer, A. and Krupinska, K. (2005) DNA-binding proteins of the Whirly family in Arabidopsis thaliana are targeted to the organelles. FEBS Lett. 579, 3707-3712.
Lai, Z., Vinod, K., Zheng, Z., Fan, B. and Chen, Z. (2008) Roles of Arabidopsis WRKY3 and WRKY4 transcription factors in plant responses to pathogens. BMC Plant Biol. 8, 68.
Li, J., Brader, G. and Palva, E.T. (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, J.F. and Wu, S.H. (2004) Molecular events in senescing Arabidopsis leaves. Plant J. 39, 612-628.
Miao, Y. and Zentgraf, U. (2007) The antagonist function of Arabidopsis WRKY53 and ESR/ESP in leaf senescence is modulated by the jasmonic and salicylic acid equilibrium. Plant Cell, 19, 819-830.
Miao, Y., Laun, T., Zimmermann, P. and Zentgraf, U. (2004) Targets of the WRKY53 transcription factor and its role during leaf senescence in Arabidopsis. Plant Mol. Biol. 55, 853-867.
Miao, Y., Laun, T., Smykowski, A. and Zentgraf, U. (2007) 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.
Miao, Y., Smykowski, A. and Zentgraf, U. (2008) A novel upstream regulator of WRKY53 transcription during leaf senescence of Arabidopsis thaliana. Plant Biol. 10(Suppl. 1), 110-120.
Peng, M.S., Hannam, C., Gu, H.L., Bi, Y.M. and Rothstein, S.J. (2007) A mutation in NLA, which encodes a RING-type ubiquitin ligase, disrupts the adaptability of Arabidopsis to nitrogen limitation. Plant J. 50, 320-337.
Pfaffl, M.W. (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29, e45.
Qiu, J.L., Fiil, B.K., Petersen, K. et al. (2008) Arabidopsis MAP kinase 4 regulates gene expression through transcription factor release in the nucleus. EMBO J. 27, 2214-2221.
Raab, S., Drechsel, G., Zarepour, M., Hartung, W., Koshiba, T., Bittner, F. and Hoth, S. (2009) Identification of a novel E3 ubiquitin ligase that is required for suppression of premature senescence in Arabidopsis. Plant J. 59, 39-51.
Robatzek, S. and Somssich, I.E. (2002) Targets of AtWRKY6 regulation during plant senescence and pathogen defense. Genes Dev. 16, 1139-1149.
Schwarz, S.E., Rosa, J.L. and Scheffner, M. (1998) Characterization of human hect domain family members and their interaction with UbcH5 and UbcH7. J. Biol. Chem. 273, 12148-12154.
Sheen, J. (2001) Signal transduction in maize and Arabidopsis mesophyll protoplasts. Plant Physiol. 127, 1466-1475.
Ülker, B. and Somssich, I.E. (2004) WRKY factors: from DNA binding towards biological function. Curr. Opin. Plant Biol. 7, 491-498.
Ülker, B., Mukhtar, M.S. and Somssich, I.E. (2007) The WRKY70 transcription factor of Arabidopsis influences both the plant senescence and defence signaling pathways. Planta, 226, 125-137.
Woo, H.R., Chung, K.M., Park, J.H., Oh, S.A., Ahn, T., Hong, S.H., Jang, S.K. and Nam, H.G. (2001) ORE9, an F-box protein that regulates leaf senescence in Arabidopsis. Plant Cell, 13, 1779-1790.
Xiang, C.B., Han, P., Lutziger, I., Wang, K. and Oliver, D.J. (1999) A mini binary vector series for plant transformation. Plant Mol. Biol. 40, 711-717.
Yang, C.W., González-Lamothe, R., Ewan, R.A., Rowland, O., Yoshioka, H., Shenton, M., Ye, H., O’Donnell, E., Jones, J.D. and Sadanandom, A. (2006) The E3 ubiquitin ligase activity of Arabidopsis PLANT U-BOX17 and its functional tobacco homolog ACRE276 are required for cell death and defense. Plant Cell, 18, 1084-1098.
Zentgraf, U., Kolb, D., Laun, T. and 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. and Zentgraf, U. (2006) Senescence specific regulation of catalases in Arabidopsis thaliana (L.) Heynh. Plant Cell Environ. 29, 1049-1060.