Targeting MAGO proteins with a peptide aptamer reinforces their essential roles in multiple rice developmental pathways
Targeting MAGO proteins with a peptide aptamer reinforces their essential roles in multiple rice developmental pathways
复制标题
用肽适体靶向 MAGO 蛋白可增强其在多种水稻发育途径中的重要作用
DOI：
10.1111/tpj.12672
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发表时间：
2014-12-01
期刊：
影响因子：
7.2
通讯作者：
He, Chaoying
中科院分区：
文献类型：
--
作者：
Gong, Pichang;Quan, Hui;He, Chaoying
肽适体是一种人工合成的短肽，可以干扰其靶蛋白的生物学功能，然而，这项技术尚未应用于植物功能基因组学。MAGO和Y14是外显子连接复合物（EJC）的两个核心亚基，在真核生物中形成专性异二聚体。水稻（Oryza sativa L.）（水稻）中，这两个基因中的每一个都有两个同源物，分别命名为OsMAGO 1和OsMAGO 2，以及OsY 14 a和OsY 14 b。在这里，我们的特点是16-氨基酸肽适体（PAP）的水稻MAGO蛋白。PAP和水稻Y14竞争性结合水稻MAGO蛋白。通过在转基因水稻中表达适体来特异性靶向MAGO蛋白并不影响MAGO蛋白的内源合成和积累;然而，在多个器官中观察到的表型变异与携带RNA干扰（RNAi）构建体的转基因水稻中MAGO和/或OsY 14 a转录本和MAGO蛋白的积累严重下调的表型变异相似。形态学上，转适体基因植株矮小，花发育异常，雄蕊的药室内壁和绒毡层的降解和吸收减少，证实了EJC核心异二聚体在水稻发育、生长和繁殖中起重要作用。本研究表明，作为RNAi的一种补充手段，肽适体是干扰高等植物蛋白质功能的有力工具。
Peptide aptamers are artificial short peptides that potentially interfere with the biological roles of their target proteins; however, this technology has not yet been applied to plant functional genomics. MAGO and Y14, the two core subunits of the exon junction complex (EJC), form obligate heterodimers in eukaryotes. In Oryza sativa L. (rice), each of the two genes has two homologs, designated OsMAGO1 and OsMAGO2, and OsY14a and OsY14b, respectively. Here, we characterized a 16-amino acida peptide aptamer (PAP) for the rice MAGO proteins. PAP and rice Y14 bound competitively to rice MAGO proteins. Specifically targeting the MAGO proteins by expressing the aptamer in transgenic rice plants did not affect the endogenous synthesis and accumulation of MAGO proteins; however, the phenotypic variations observed in multiple organs phenocopied those of transgenic rice plants harboring RNA interference (RNAi) constructs in which the accumulation of MAGO and/or OsY14a transcripts and MAGO proteins was downregulated severely. Morphologically, the aptamer transgenic plants were short with abnormally developed flowers, and the stamens exhibited reduced degradation and absorption of both the endothecium and tapetum, thus confirming that EJC core heterodimers play essential roles in rice development, growth and reproduction. This study reveals that as a complementary approach of RNAi, peptide aptamers are powerful tools for interfering with the function of proteins in higher plants.