Bai Ming-Yi, Zhang Li-Ying, Gampala Srinivas S, Zhu Sheng-Wei, Song Wen-Yuan, Chong Kang, Wang Zhi-Yong
Key Laboratory of Photosynthesis and Environmental Molecular Biology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Proc Natl Acad Sci U S A. 2007 Aug 21;104(34):13839-44. doi: 10.1073/pnas.0706386104. Epub 2007 Aug 15.
Brassinosteroids (BR) are essential growth hormones found throughout the plant kingdom. BR bind to the receptor kinase BRI1 on the cell surface to activate a signal transduction pathway that regulates nuclear gene expression and plant growth. To understand the downstream BR signaling mechanism in rice, we studied the function of OsBZR1 using reverse genetic approaches and identified OsBZR1-interacting proteins. Suppressing OsBZR1 expression by RNAi resulted in dwarfism, erect leaves, reduced BR sensitivity, and altered BR-responsive gene expression in transgenic rice plants, demonstrating an essential role of OsBZR1 in BR responses in rice. Moreover, a yeast two-hybrid screen identified 14-3-3 proteins as OsBZR1-interacting proteins. Mutation of a putative 14-3-3-binding site of OsBZR1 abolished its interaction with the 14-3-3 proteins in yeast and in vivo. Such mutant OsBZR1 proteins suppressed the phenotypes of the Arabidopsis bri1-5 mutant and showed an increased nuclear distribution compared with the wild-type protein, suggesting that 14-3-3 proteins directly inhibit OsBZR1 function at least in part by reducing its nuclear localization. These results demonstrate a conserved function of OsBZR1 and an important role of 14-3-3 proteins in brassinosteroid signal transduction in rice.
油菜素甾醇(BR)是在整个植物界都存在的必需生长激素。BR与细胞表面的受体激酶BRI1结合,以激活一条调节核基因表达和植物生长的信号转导途径。为了了解水稻中BR信号转导的下游机制,我们采用反向遗传学方法研究了OsBZR1的功能,并鉴定了与OsBZR1相互作用的蛋白。通过RNA干扰抑制OsBZR1的表达,导致转基因水稻植株出现矮化、叶片直立、BR敏感性降低以及BR响应基因表达改变的现象,这表明OsBZR1在水稻的BR反应中起着至关重要的作用。此外,酵母双杂交筛选鉴定出14-3-3蛋白是与OsBZR1相互作用的蛋白。OsBZR1一个假定的14-3-3结合位点发生突变后,在酵母和体内均消除了它与14-3-3蛋白的相互作用。与野生型蛋白相比,这种突变的OsBZR1蛋白抑制了拟南芥bri1-5突变体的表型,并显示出核分布增加,这表明14-3-3蛋白至少部分地通过减少OsBZR1的核定位来直接抑制其功能。这些结果证明了OsBZR1的保守功能以及14-3-3蛋白在水稻油菜素甾醇信号转导中的重要作用。