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Nucleic Acids Research Advance Access published online on May 7, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm273
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Pathogen-induced binding of the soybean zinc finger homeodomain proteins GmZF-HD1 and GmZF-HD2 to two repeats of ATTA homeodomain binding site in the calmodulin isoform 4 (GmCaM4) promoter

Hyeong Cheol Park1,2, Man Lyang Kim1, Sang Min Lee1,2, Jeong Dong Bahk1,2, Dae-Jin Yun1,2, Chae Oh Lim1,2, Jong Chan Hong1,2, Sang Yeol Lee1,2, Moo Je Cho1 and Woo Sik Chung1,2,*

1Division of Applied Life Science (BK21 program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, 660-701, Korea and 2Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju, 660-701, Korea

*To whom correspondence should be addressed. Tel: +82 55 751 6254; Fax: +82 55 759 9363; Email: chungws{at}gnu.ac.kr Correspondence may also be addressed to Moo Je Cho. Tel: +82-55-751-6254; Fax: +82-55-759-9363; Email: choslab{at}gnu.ac.kr

Received December 1, 2006. Revised April 10, 2007. Accepted April 10, 2007.

Calmodulin (CaM) is involved in defense responses in plants. In soybean (Glycine max), transcription of calmodulin isoform 4 (GmCaM4) is rapidly induced within 30 min after pathogen stimulation, but regulation of the GmCaM4 gene in response to pathogen is poorly understood. Here, we used the yeast one-hybrid system to isolate two cDNA clones encoding proteins that bind to a 30-nt A/T-rich sequence in the GmCaM4 promoter, a region that contains two repeats of a conserved homeodomain binding site, ATTA. The two proteins, GmZF-HD1 and GmZF-HD2, belong to the zinc finger homeodomain (ZF-HD) transcription factor family. Domain deletion analysis showed that a homeodomain motif can bind to the 30-nt GmCaM4 promoter sequence, whereas the two zinc finger domains cannot. Critically, the formation of super-shifted complexes by an anti-GmZF-HD1 antibody incubated with nuclear extracts from pathogen-treated cells suggests that the interaction between GmZF-HD1 and two homeodomain binding site repeats is regulated by pathogen stimulation. Finally, a transient expression assay with Arabidopsis protoplasts confirmed that GmZF-HD1 can activate the expression of GmCaM4 by specifically interacting with the two repeats. These results suggest that the GmZF-HD1 and –2 proteins function as ZF-HD transcription factors to activate GmCaM4 gene expression in response to pathogen.

Present addresses: Hyeong Cheol Park, Center for Plant Environmental Stress Physiology, Department of Horticulture & Landscape Architecture, Purdue University, West Lafayette, Indiana 47907, USA Man Lyang Kim, Department of Infection Biology, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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