Mechanism of phytochrome signaling mediated by protein interaction

蛋白质相互作用介导的光敏色素信号传导机制

基本信息

批准号：
16580076
负责人：
KOHCHI Takayuki
金额：
$ 2.37万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580076/
关键词：
photoreceptor phytochrome chromophore photomorphogenesis Arabidopsis thaliana protein interaction phytochromobilin 暗反転

项目摘要

Sessile and photoautotrophic plants have developed very sensitive and sophisticated system to recognize light conditions properly for their survival and successful propagation. Phytochrome is a major photoreceptor in plants. However, the mechanisms of signal transduction mediated by phytochrome is largely unknown. The protein interaction should be a key mechanism for signaling for phytochrome since phytochrome have very specific character showing photoreversible conformation changes by illumination. To study protein interaction with phytochrome, the photoactive phytochrome with chromophore is required. By co-expression of heme oxygenase and various bilin reductase(s) in a single operon in conjunction with apophytochrome using two compatible plasmids, we developed a system to produce phytochromes with various chromophores in Escherichia coli. The metabolic engineering of bacteria for the production of various bilins for assembly into phytochromes will facilitate the molecular analysis of photoreceptors. We also generated transgenic plants that express epitope-tagged phytochromes. The introduced fusion genes were active and functional to rescue phytochrome mutant phenotype of phyA and phyB. Another approach we took to identify interacting proteins was yeast two-hybrid assays. A novel transcription factor containing zinc finger domain was identified. The specificity of interaction for wavelength was verified by using expressed proteins in vitro. The identified protein is a good candidate to mediated photobiological response since the knock-out mutant showed delayed flowering phenotype.In addition to Arabidopsis works, we isolated several genes and corresponding mutants for heme oxygenase and phytochromobilin synthase for phytocrome chromophore biosynthesis from maize, pea and tomato. In particular, the tomato AUREA protein was expressed at higher levels in soluble form in E.coli.

无梗和光自营养植物已经开发出非常敏感和复杂的系统，以识别其生存和成功传播的光条件。植物色素是植物中的主要光感受器。但是，植物色素介导的信号转导机制在很大程度上尚不清楚。蛋白质相互作用应该是植物色素信号传导的关键机制，因为植物色素具有非常特异性的特征，显示出可爱的构象通过照明而变化。为了研究与植物色素的蛋白质相互作用，需要光活性植物色素与发色团。通过使用两种兼容质粒的单一操纵子中的血红素氧酶和各种bilin还原酶的共表达，并结合脂肪色素，我们开发了一个系统，可以在大肠杆菌中生产具有各种发色团的植物色素。细菌用于生产各种bilins以组装成植物色素的代谢工程将促进感光体的分子分析。我们还产生了表达表现标记的植物色素的转基因植物。引入的融合基因具有活性和功能性，可挽救Phya和Phyb的植物色素突变体表型。我们采用的另一种方法是鉴定相互作用的蛋白质是酵母双杂交测定法。鉴定出一种含有锌指域的新型转录因子。通过在体外使用表达的蛋白质来验证相互作用的特异性。鉴定蛋白是介导的光生物学反应的良好候选者，因为敲除突变体显示出延迟的开花表型。在拟南芥作品中，我们分离了几种基因，并分离出植物蛋白酶氧化物酶和植物色素蛋白同步的相应突变体，用于植物生物团生物合成生物合成。特别是，西红柿耳蛋白以较高的水平以可溶性形式表达。

项目成果

期刊论文数量（38）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Isolation and functional characterization of fatty acid D5-elongase gene from the liverwort Marchantia polymorpha L.

地钱地钱脂肪酸D5-延长酶基因的分离及功能鉴定。

DOI：
发表时间：
2006
期刊：
FEBS Lett. 580
影响因子：
0
作者：
Kajikawa;M.;Yamato;K.T.;Sakai;Y.;Fukuzawa;H.;Ohyama;K.;Kohchi;T.
通讯作者：
T.

The tomato photomorphogenetic mutant, aurea, is deficient in phytocbromobilin synthase for phytochrome chromophore biosynthesis.

番茄光形态发生突变体 aurea 缺乏用于光敏色素发色团生物合成的植物动素合酶。

DOI：
发表时间：
2005
期刊：
Plant Cell Physiol. 46
影响因子：
0
作者：
Muramoto;T.;Kami;C.;Kataoka;H.;Iwata;N.;Linley;P.J.;Mukougawa;K.;Yokota;A.;Kohchi;T.
通讯作者：
T.

The Elml (ZmHy2) Gene of Maize Encodes a Phytochromobilin Synthase.

玉米的 Elml (ZmHy2) 基因编码植物色素合酶。

DOI：
发表时间：
2004
期刊：
Plant Physiology 136
影响因子：
0
作者：
Sawers;R.J.;Linley;P.J.;Gutierrez-Marcos;J.F.;Delli-Bovi;T.;Farmer;P.R.;Kohchi;T.;Terry;M.J.;Brutnell;T.
通讯作者：
T.

Molecular Characterization of the Cytoplasmic Interacting Protein of the Receptor Kinase IRK Expressed in the Inflorescence and Root Apices of Arabidopsis