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MPK4 Phosphorylation Dynamics and Interacting Proteins in Plant Immunity.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-01-28 , DOI: 10.1021/acs.jproteome.8b00345 Tong Zhang , Jacqueline D Schneider , Chuwei Lin , Sisi Geng , Tianyi Ma , Sheldon R Lawrence , Craig P Dufresne 1 , Alice C Harmon , Sixue Chen
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-01-28 , DOI: 10.1021/acs.jproteome.8b00345 Tong Zhang , Jacqueline D Schneider , Chuwei Lin , Sisi Geng , Tianyi Ma , Sheldon R Lawrence , Craig P Dufresne 1 , Alice C Harmon , Sixue Chen
Affiliation
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Arabidopsis MAP kinase 4 (MPK4) has been proposed to be a negative player in plant immunity, and it is also activated by pathogen-associated molecular patterns (PAMPs), such as flg22. The molecular mechanisms by which MPK4 is activated and regulates plant defense remain elusive. In this study, we investigated Arabidopsis defense against a bacterial pathogen Pseudomonas syringae pv tomato ( Pst) DC3000 when Brassica napus MPK4 ( BnMPK4) is overexpressed. We showed an increase in pathogen resistance and suppression of jasmonic acid (JA) signaling in the BnMPK4 overexpressing (OE) plants. We also showed that the OE plants have increased sensitivity to flg22-triggered reactive oxygen species (ROS) burst in guard cells, which resulted in enhanced stomatal closure compared to wild-type (WT). During flg22 activation, dynamic phosphorylation events within and outside of the conserved TEY activation loop were observed. To elucidate how BnMPK4 functions during the defense response, we used immunoprecipitation coupled with mass spectrometry (IP-MS) to identify BnMPK4 interacting proteins in the absence and presence of flg22. Quantitative proteomic analysis revealed a shift in the MPK4-associated protein network, providing insight into the molecular functions of MPK4 at the systems level.
中文翻译：
植物免疫中的MPK4磷酸化动力学和相互作用蛋白。
拟南芥MAP激酶4（MPK4）被认为是植物免疫力的消极参与者，并且也被病原体相关分子模式（PAMP）（例如flg22）激活。激活MPK4并调节植物防御的分子机制仍然难以捉摸。在这项研究中，我们调查了甘蓝型油菜MPK4（BnMPK4）过表达时拟南芥对细菌病原体丁香假单胞菌pv番茄（Pst）DC3000的防御作用。我们显示在BnMPK4过表达（OE）植物中病原体抗性的增加和茉莉酸（JA）信号的抑制。我们还显示，OE植物对flg22触发的保卫细胞中触发的活性氧（ROS）爆发具有更高的敏感性，与野生型（WT）相比，可增强气孔关闭。在激活flg22期间，观察到保守的TEY激活环内外的动态磷酸化事件。为了阐明BnMPK4在防御反应中的功能，我们使用免疫沉淀结合质谱（IP-MS）来鉴定在不存在flg22的情况下BnMPK4相互作用的蛋白。定量蛋白质组学分析揭示了与MPK4相关的蛋白质网络的转变，从而洞察了MPK4在系统水平上的分子功能。
更新日期：2019-02-07
中文翻译：
植物免疫中的MPK4磷酸化动力学和相互作用蛋白。
拟南芥MAP激酶4（MPK4）被认为是植物免疫力的消极参与者，并且也被病原体相关分子模式（PAMP）（例如flg22）激活。激活MPK4并调节植物防御的分子机制仍然难以捉摸。在这项研究中，我们调查了甘蓝型油菜MPK4（BnMPK4）过表达时拟南芥对细菌病原体丁香假单胞菌pv番茄（Pst）DC3000的防御作用。我们显示在BnMPK4过表达（OE）植物中病原体抗性的增加和茉莉酸（JA）信号的抑制。我们还显示，OE植物对flg22触发的保卫细胞中触发的活性氧（ROS）爆发具有更高的敏感性，与野生型（WT）相比，可增强气孔关闭。在激活flg22期间，观察到保守的TEY激活环内外的动态磷酸化事件。为了阐明BnMPK4在防御反应中的功能，我们使用免疫沉淀结合质谱（IP-MS）来鉴定在不存在flg22的情况下BnMPK4相互作用的蛋白。定量蛋白质组学分析揭示了与MPK4相关的蛋白质网络的转变，从而洞察了MPK4在系统水平上的分子功能。
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