Kawamura Masakazu, Ito Shogo, Nakamichi Norihito, Yamashino Takafumi, Mizuno Takeshi
Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan.
Biosci Biotechnol Biochem. 2008 May;72(5):1307-16. doi: 10.1271/bbb.70804. Epub 2008 May 7.
In the model higher plant Arabidopsis thaliana, the CCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) gene plays important circadian clock-associated roles. The CCA1 protein is a member of a small subfamily of single MYB-related transcription factors. This family consists of several homologous CCA1-like transcription factors, including the closest homolog LHY (LATE ELONGATED HYPOCOTYL). To gain insight into the molecular function of CCA1 and its homologs, here we took a unique genetic approach that was recently developed for Arabidopsis thaliana. Through this strategy, referred to as CRES-T (Chimeric REpressor Silencing Technology), a transgenic plant was constructed to produce a dominant negative transcriptional repressor (designated CCA1-SRDX). By employing the resulting transgenic lines, together with previously established cca1 lhy double mutant and CCA1-ox (over-expressing) plants, their circadian clock-associated phenotypes were examined and compared with each other. The observed clock-associated phenotypes of the CCA1-SRDX plants were very similar to those of CCA1-ox, but not to those of cca1 lhy, suggesting that CCA1 acts predominantly as a transcriptional repressor in nature. However, the developmental morphology (or architecture) of adult CCA1-SRDX plants were quite different from that of CCA1-ox, suggesting that CCA1 might also be implicated, directly or indirectly, in an as yet unknown circadian-associated output pathway at a late developmental stage.
在模式高等植物拟南芥中,CCA1(生物钟相关1)基因发挥着重要的生物钟相关作用。CCA1蛋白是单个MYB相关转录因子小亚家族的成员。该家族由几个同源的CCA1样转录因子组成,包括最接近的同源物LHY(晚伸长下胚轴)。为了深入了解CCA1及其同源物的分子功能,我们采用了一种最近为拟南芥开发的独特遗传方法。通过这种称为CRES-T(嵌合阻遏物沉默技术)的策略,构建了一种转基因植物以产生显性负转录阻遏物(命名为CCA1-SRDX)。通过使用所得的转基因株系,连同先前建立的cca1 lhy双突变体和CCA1-ox(过表达)植物,研究并比较了它们的生物钟相关表型。观察到的CCA1-SRDX植物的生物钟相关表型与CCA1-ox的非常相似,但与cca1 lhy的不同,这表明CCA1在本质上主要作为转录阻遏物起作用。然而,成年CCA1-SRDX植物的发育形态(或结构)与CCA1-ox的有很大不同,这表明CCA1在发育后期可能也直接或间接地参与了一个未知的生物钟相关输出途径。