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ENVIRONMENTAL STRESS AND ADAPTATION

Functional Analysis of Arabidopsis Ethylene-Responsive Element Binding Protein Conferring Resistance to Bax and Abiotic Stress-Induced Plant Cell Death¹

Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo 113–0032, Japan (T.O., L.P., M.K.-Y., L.-H.Y., H.U.); Iwate Biotechnology Research Center, Kitakami, Iwate 024–0003, Japan (S.Y., H.U.); Gene Function Research Laboratory, National Institute of Advanced Industrial Science and Technology, Tsukuba 305–8562, Japan (T.K., M.O.-T.); and Department of Applied Life Sciences, Kyoto University, Kitashirakawa, Sakyo, Kyoto 606–8502, Japan (S.K., F.S.)

Arabidopsis (Arabidopsis thaliana) ethylene-responsive element binding protein (AtEBP) gene was isolated as a suppressor of Bax-induced cell death by functional screening in yeast (Saccharomyces cerevisiae). To further examine the cell death suppressive action of AtEBP in plant cells, we established transgenic tobacco (Nicotiana tabacum) plants overexpressing AtEBP as well as transgenic tobacco plants ectopically expressing mouse Bax protein under a dexamethasone-inducible promoter. We prepared the crosses of the selective lines of each transgenic plant, which were evaluated in terms of cell death suppression activity. Results indicate that AtEBP suppressed Bax-induced cell death in tobacco plants, an action also associated with a lowered level of ion leakage. Furthermore, tobacco Bright Yellow-2 cells overexpressing AtEBP conferred resistance to hydrogen peroxide (H₂O₂) and heat treatments. AtEBP protein localized in the nucleus and functioned as an in vivo transcription activator as confirmed in transient assays and experiments using stable transgenic system. Up-regulation of defense genes was observed in transgenic Arabidopsis plants overexpressing AtEBP. Based on the analysis of mRNA accumulation in ethylene-related mutants, the position of AtEBP in signaling pathway is presented.

² Present address: Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki Sakyo-ku, Kyoto 606–8585, Japan.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.063586.

^* Corresponding author; e-mail uchimiya{at}iam.u-tokyo.ac.jp; fax 81–3–5841–8466.

Received March 30, 2005; returned for revision March 30, 2005; accepted April 13, 2005.

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