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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Cell Death Suppressor Arabidopsis Bax Inhibitor-1 Is Associated with Calmodulin Binding and Ion Homeostasis^1,[OA]

Institute of Molecular and Cellular Biosciences, The University of Tokyo, Bunkyo-ku, Tokyo 113–0032, Japan (Y.I.-O., M.N., H.U., M.K.-Y.); Nagoya University Bioscience Center, Nagoya University, Chikusa, Nagoya 464–8601, Japan (S.M.); Iwate Biotechnology Research Center, Kitakami, Iwate 024–0003, Japan (H.U.); and Japan Science and Technology Agency, Core Research for Evolutional Science and Technology, Kawaguchi, Saitama 332–0012, Japan (M.K.-Y.)

Cell death suppressor Bax inhibitor-1 (BI-1), an endoplasmic reticulum membrane protein, exists in a wide range of organisms. The split-ubiquitin system, overlay assay, and bimolecular fluorescence complementation analysis demonstrated that Arabidopsis (Arabidopsis thaliana) BI-1 (AtBI-1) interacted with calmodulin in yeast (Saccharomyces cerevisiae) and in plant cells. Furthermore, AtBI-1 failed to rescue yeast mutants lacking Ca²⁺ ATPase (Pmr1 or Spf1) from Bax-induced cell death. Pmr1 and Spf1, p-type ATPases localized at the inner membrane, are believed to be involved in transmembrane movement of calcium ions in yeast. Thus, the presence of intact Ca²⁺ ATPases was essential for AtBI-1-mediated cell death suppression in yeast. To investigate the effect of AtBI-1 on calcium homeostasis, we evaluated sensitivity against cyclopiazonic acid (CPA), an inhibitor of sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase in AtBI-1-overexpressing or knock-down transgenic Arabidopsis plants. These plants demonstrated altered CPA or ion stress sensitivity. Furthermore, AtBI-1-overexpressing cells demonstrated an attenuated rise in cytosolic calcium following CPA or H₂O₂ treatment, suggesting that AtBI-1 affects ion homeostasis in plant cell death regulation.

² This paper is dedicated to the memory of the late Prof. Muto.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Maki Kawai-Yamada (mkawai{at}iam.u-tokyo.ac.jp).

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www.plantphysiol.org/cgi/doi/10.1104/pp.106.090878

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

Received October 5, 2006; accepted November 21, 2006; published December 1, 2006.

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