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First published online November 17, 2004; 10.1105/tpc.104.026112

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The Plant Cell 16:3448-3459 (2004)
© 2004 American Society of Plant Biologists

Solution Structure of the B3 DNA Binding Domain of the Arabidopsis Cold-Responsive Transcription Factor RAV1{boxw}

Kazuhiko Yamasakia,b,1, Takanori Kigawab, Makoto Inoueb, Masaru Tatenoa,c, Tomoko Yamasakia, Takashi Yabukib, Masaaki Aokib, Eiko Sekib, Takayoshi Matsudab, Yasuko Tomob, Nobuhiro Hayamib, Takaho Teradab,d, Mikako Shirouzub,d, Takashi Osanaib, Akiko Tanakab, Motoaki Sekie,f, Kazuo Shinozakie,f,g and Shigeyuki Yokoyamab,d,h

a Age Dimension Research Center, National Institute of Advanced Industrial Science and Technology, Tsukuba 305-8566, Japan
b Protein Research Group, RIKEN Genomic Sciences Center, Yokohama 230-0045, Japan
c Center for Biological Resources and Informatics, Tokyo Institute of Technology, Yokohama 226-8501, Japan
d RIKEN Harima Institute at SPring8, Hyogo 679-5148, Japan
e Laboratory of Plant Molecular Biology, RIKEN Tsukuba Institute, Tsukuba 305-0074, Japan
f Plant Functional Genomics Research Group, RIKEN Genomic Sciences Center,Yokohama 230-0045, Japan
g Institute of Biological Sciences, University of Tsukuba, Tsukuba 305-8572, Japan
h Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan

1 To whom correspondence should be addressed. E-mail k-yamasaki{at}aist.go.jp; fax 81-298-61-2703.

The B3 DNA binding domain is shared amongst various plant-specific transcription factors, including factors involved in auxin-regulated and abscisic acid–regulated transcription. Herein, we report the NMR solution structure of the B3 domain of the Arabidopsis thaliana cold-responsive transcription factor RAV1. The structure consists of a seven-stranded open ß-barrel and two {alpha}-helices located at the ends of the barrel and is significantly similar to the structure of the noncatalytic DNA binding domain of the restriction enzyme EcoRII. An NMR titration experiment revealed a DNA recognition interface that enabled us to propose a structural model of the protein–DNA complex. The locations of the DNA-contacting residues are also likely to be similar to those of the EcoRII DNA binding domain.




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