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Published on August 27, 2004; 10.1104/pp.104.046599

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Received May 17, 2004
Returned for revision May 31, 2004
Accepted June 1, 2004

Arabidopsis Cys2/His2-Type Zinc-Finger Proteins Function as Transcription Repressors under Drought, Cold, and High-Salinity Stress Conditions

Hideki Sakamoto , Kyonoshin Maruyama , Yoh Sakuma , Tetsuo Meshi , Masaki Iwabuchi , Kazuo Shinozaki , and Kazuko Yamaguchi-Shinozaki *

Biological Resources Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan (H.S., K.M., Y.S., K.Y.-S.); Department of Botany, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan (H.S., T.M.); Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Institute of Physical and Chemical Research, Tsukuba, Ibaraki 305-0074, Japan (K.S.); Research Institute for Biological Sciences, Kayo-cho, Okayama 716-1241, Japan (M.I.); National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan (T.M., M.I.); Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan (K.Y.-S.); and Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Kawaguchi-shi, Saitama 332-0012, Japan (K.S., K.Y.-S.)

* Corresponding author; email: kazukoys{at}jircas.affrc.go.jp.

ZPT2-related proteins that have two canonical Cys-2/His-2-type zinc-finger motifs in their molecules are members of a family of plant transcription factors. To characterize the role of this type of protein, we analyzed the function of Arabidopsis L. Heynh. genes encoding four different ZPT2-related proteins (AZF1, AZF2, AZF3, and STZ). Gel-shift analysis showed that the AZFs and STZ bind to A(G/C)T repeats within an EP2 sequence, known as a target sequence of some petunia (Petunia hybrida) ZPT2 proteins. Transient expression analysis using synthetic green fluorescent protein fusion genes indicated that the AZFs and STZ are preferentially localized to the nucleus. These four ZPT2-related proteins were shown to act as transcriptional repressors that down-regulate the transactivation activity of other transcription factors. RNA gel-blot analysis showed that expression of AZF2 and STZ was strongly induced by dehydration, high-salt and cold stresses, and abscisic acid treatment. Histochemical analysis of {beta}-glucuronidase activities driven by the AZF2 or STZ promoters revealed that both genes are induced in leaves rather than roots of rosette plants by the stresses. Transgenic Arabidopsis overexpressing STZ showed growth retardation and tolerance to drought stress. These results suggest that AZF2 and STZ function as transcriptional repressors to increase stress tolerance following growth retardation.




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