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JXB Advance Access published online on March 3, 2003
Journal of Experimental Botany, doi:10.1093/jxb/erg136
© 2003 by Oxford University Press
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Received July 15, 2002; accepted December 17, 2002
© 2003 Society for Experimental Biology

RESEARCH PAPER

Isolation of tobacco ubiquitin-conjugating enzyme cDNA in a yeast two-hybrid system with tobacco ERF3 as bait and its characterization of specific interaction

Tomotsugu Koyama 1, Takashi Okada 2, Sakihito Kitajima 2, Masaru Ohme-Takagi 3, Hideaki Shinshi 4, Fumihiko Sato 1*

1 Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan; Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan
2 Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
3 Gene Function Research Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1 Higashi, Tsukuba 305-0046, Japan
4 Institute of Molecular and Cell Biology, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1 Higashi, Tsukuba 305-0046, Japan

* To whom correspondence should be addressed. E-mail: fumihiko{at}kais.kyoto-u.ac.jp.


  Abstract

Tobacco ETHYLENE-RESPONSIVE FACTOR3 (ERF3) is a member of the ERF-domain transcription factors and has a transcriptional repressor activity, whereas other ERF proteins show activation activity. To understand the regulation of ERF3-repressor activity, protein(s) were screened which interact with ERF3 in a yeast two-hybrid system. A partial sequence (B8) of NtUBC2, a tobacco ubiquitin-conjugating enzyme was isolated. This B8 specifically interacted with ERF3 in the yeast two-hybrid system. Further analyses revealed that the region unique to ERF3 interacted with B8. The physiological functions of NtUBC2 and the stability of ERF3 are discussed in relation to the regulation of the repression activity of ERF3.

Key words: Degradation, ethylene responsive factor (ERF), repressor, transient assay, two-hybrid system, ubiquitin-conjugating enzyme (UBC).


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