Plant Physiology Preview Published on May 13, 2005; 10.1104/pp.104.057752
Received December 7, 2004
Returned for revision January 18, 2005
Accepted January 21, 2005
Analysis of a Sugar Response Mutant of Arabidopsis Identified a Novel B3 Domain Protein That Functions as an Active Transcriptional Repressor
Hironaka Tsukagoshi *, Takanori Saijo , Daisuke Shibata , Atsushi Morikami , and Kenzo Nakamura
Laboratory of Biochemistry, Graduate School of Bioagricultural Science, Nagoya University, Chikusa, Nagoya 464-8601, Japan
Kazusa DNA Research Institute, Kisarazu, Chiba 292-0818, Japan
College of Bioscience and Biotechnology, Chubu University, Kasugai, Aichi 487-8501, Japan
* Corresponding author; email: i032011d{at}mbox.nagoya-u.ac.jp.
A recessive mutation hsi2 of Arabidopsis (Arabidopsis thaliana) expressing luciferase (LUC) under control of a short promoter derived from a sweet potato (Ipomoea batatas) sporamin gene (Spomin::LUC) caused enhanced LUC expression under both low- and high-sugar conditions, which was not due to increased level of abscisic acid. The hsi2 mutant contained a nonsense mutation in a gene encoding a protein with B3 DNA-binding domain. HSI2 and two other Arabidopsis proteins appear to constitute a novel subfamily of B3 domain proteins distinct from ABI3, FUS3, and LEC2, which are transcription activators involved in seed development. The C-terminal part of HSI2 subfamily proteins contained a sequence similar to the ERF-associated amphiphilic repression (EAR) motif. Deletion of the C-terminal portion of HSI2 lost in the hsi2 mutant caused reduced nuclear targeting of HSI2. Null allele of HSI2 showed even higher Spomin::LUC expression than the hsi2 mutant, whereas overexpression of HSI2 reduced the LUC expression. Transient coexpression of 35S::HSI2 with Spomin::LUC in protoplasts repressed the expression of LUC activity, and deletion or mutation of the EAR motif significantly reduced the repression activity of HSI2. These results indicate that HSI2 and related proteins are B3 domain-EAR motif active transcription repressors.
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