A proteomics study of brassinosteroid response in Arabidopsis
A proteomics study of brassinosteroid response in Arabidopsis
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DOI：
10.1074/mcp.m700123-mcp200
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发表时间：
2007-12-01
影响因子：
7
通讯作者：
Wang, Zhi-Yong
中科院分区：
文献类型：
--
作者：
Deng, Zhiping;Zhang, Xin;Wang, Zhi-Yong
植物类固醇激素油菜素类固醇(BRs)在广泛的发育和生理过程中发挥着重要作用。BR信号如何调控不同的过程仍不清楚。为了了解BR反应的分子细节，我们利用二维DGE结合LC-MS/MS对拟南芥中BR调节蛋白进行了蛋白质组学研究。我们鉴定了42个BR调节蛋白，它们被预测在BR调节特定的细胞过程中发挥潜在作用，如信号传递、细胞骨架重排、小泡运输以及激素和维生素的生物合成。对BR不敏感突变体BRI1-116和BR超敏突变体BZR1-1D的分析表明，5种蛋白质(PATL1、PATL2、THI1、AtMDAR3和NADP-ME2)在BR处理和突变体中都有影响，表明它们在BR作用中具有重要作用。用插入敲除突变体或免疫印迹对选定的蛋白质进行进一步研究。有趣的是，在以前的微阵列研究中，大约80%的BR反应蛋白没有被识别出来，并且在BR突变体中蛋白质和RNA变化之间的直接比较显示出非常弱的相关性。对选定基因的RT-PCR分析揭示了RNA和蛋白质反应之间的基因特异性动力学关系。此外，BR调控的BiP2蛋白的翻译后修饰在二维DGE中被检测到为斑点移位。这项研究为将BR信号与特定的细胞和生理反应联系起来的分子网络提供了新的见解。
The plant steroid hormones brassinosteroids (BRs) play an important role in a wide range of developmental and physiological processes. How BR signaling regulates diverse processes remains unclear. To understand the molecular details of BR responses, we performed a proteomics study of BR-regulated proteins in Arabidopsis using two-dimensional DIGE coupled with LC-MS/MS. We identified 42 BR-regulated proteins, which are predicted to play potential roles in BR regulation of specific cellular processes, such as signaling, cytoskeleton rearrangement, vesicle trafficking, and biosynthesis of hormones and vitamins. Analyses of the BR-insensitive mutant bri1-116 and BR-hypersensitive mutant bzr1-1D identified five proteins (PATL1, PATL2, THI1, AtMDAR3, and NADP-ME2) affected both by BR treatment and in the mutants, suggesting their importance in BR action. Selected proteins were further studied using insertion knock-out mutants or immunoblotting. Interestingly about 80% of the BR-responsive proteins were not identified in previous microarray studies, and direct comparison between protein and RNA changes in BR mutants revealed a very weak correlation. RT-PCR analysis of selected genes revealed gene-specific kinetic relationships between RNA and protein responses. Furthermore BR-regulated posttranslational modification of BiP2 protein was detected as spot shifts in two-dimensional DIGE. This study provides novel insights into the molecular networks that link BR signaling to specific cellular and physiological responses.