Quantitative and Functional Phosphoproteomic Analysis Reveals that Ethylene Regulates Water Transport via the C-Terminal Phosphorylation of Aquaporin PIP2; 1 in Arabidopsis
Quantitative and Functional Phosphoproteomic Analysis Reveals that Ethylene Regulates Water Transport via the C-Terminal Phosphorylation of Aquaporin PIP2; 1 in Arabidopsis
复制标题
定量和功能磷酸化蛋白质组学分析表明，乙烯通过拟南芥中水通道蛋白 PIP2;1 的 C 末端磷酸化来调节水运输。
DOI：
10.1016/j.molp.2015.10.001
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发表时间：
2016-01-04
期刊：
影响因子：
27.5
通讯作者：
Li, Ning
中科院分区：
文献类型：
--
作者：
Qing, Dongjin;Yang, Zhu;Li, Ning
乙烯参与调节许多细胞事件和生物过程，包括叶片和花瓣萎蔫期间的水分损失。不同的乙烯反应可以通过蛋白质磷酸化/去磷酸化和泛素/26 S蛋白酶体介导的蛋白质降解和蛋白酶切割之间的动态相互作用来调节。为了解决乙烯如何通过多分叉信号通路改变蛋白质磷酸化，我们进行了N-15稳定同位素标记为基础的，差分和定量磷酸化蛋白质组学研究空气和乙烯处理的乙烯不敏感的拟南芥双功能丧失突变体ein 3 -1/eil 1 -1。在535个非冗余的磷酸肽鉴定，两个和四个磷酸肽被上调和下调乙烯，分别。乙烯调控的水通道蛋白PIP 2; 1磷酸化与叶片水分通量和失水量呈正相关。对表达野生型和S280 A/S283 A突变的PIP 2的转基因植物进行遗传研究，并结合定量蛋白质组学、免疫印迹分析、原生质体膨胀/收缩实验和叶片失水测定;在Col-0和ein 3eil 1遗传背景下，乙烯通过增强拟南芥细胞的S280/S283在PIP 2的C末端的磷酸化; 1.未知的激酶和/或磷酸酶活性可能独立于EIN 3/EIL 1的细胞功能参与初始上调。这一发现有助于我们了解乙烯调节的叶片萎蔫，这是常见的收获后储存的植物器官。
Ethylene participates in the regulation of numerous cellular events and biological processes, including water loss, during leaf and flower petal wilting. The diverse ethylene responses may be regulated via dynamic interplays between protein phosphorylation/dephosphorylation and ubiquitin/26S proteasome-mediated protein degradation and protease cleavage. To address how ethylene alters protein phosphorylation through multi-furcated signaling pathways, we performed a N-15 stable isotope labelling-based, differential, and quantitative phosphoproteomics study on air-and ethylene-treated ethylene-insensitive Arabidopsis double loss-of-function mutant ein3-1/eil1-1. Among 535 non-redundant phosphopeptides identified, two and four phosphopeptides were up-and downregulated by ethylene, respectively. Ethylene-regulated phosphorylation of aquaporin PIP2; 1 is positively correlated with the water flux rate and water loss in leaf. Genetic studies in combination with quantitative proteomics, immunoblot analysis, protoplast swelling/shrinking experiments, and leaf water loss assays on the transgenic plants expressing both the wild-type and S280A/S283A-mutated PIP2; 1 in the both Col-0 and ein3eil1 genetic backgrounds suggest that ethylene increases water transport rate in Arabidopsis cells by enhancing S280/S283 phosphorylation at the C terminus of PIP2; 1. Unknown kinase and/or phosphatase activities may participate in the initial up-regulation independent of the cellular functions of EIN3/EIL1. This finding contributes to our understanding of ethylene-regulated leaf wilting that is commonly observed during post-harvest storage of plant organs.