Identification and functional analysis of phosphorylation residues of the Arabidopsis BOTRYTIS-INDUCED KINASE1
Identification and functional analysis of phosphorylation residues of the Arabidopsis BOTRYTIS-INDUCED KINASE1
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拟南芥BOTRYTIS诱导激酶1磷酸化残基的鉴定及功能分析
DOI：
10.1007/s13238-013-3053-6
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发表时间：
2013-09
期刊：
影响因子：
--
通讯作者：
Wenqing Shui
中科院分区：
文献类型：
--
作者：
Honggang Zhou;Cheng Yang;Zhiyong Lou;Wenqing Shui
拟南芥肉毒素诱导激酶1（BIK 1）是一种受体样细胞质激酶，在植物生长和先天免疫的多个信号通路中起重要作用。已知与细胞表面受体FLS 2和共受体激酶BAK 1形成信号传导复合物，以在感知病原体相关分子模式（PAMP）时阻断信号。虽然推测位点特异性磷酸化介导BIK 1的激活和功能，但很少有研究致力于完整分析BIK 1磷酸化残基。在这里，我们确定了19个体外自磷酸化位点的BIK 1，包括三个磷酸酪氨酸位点，从而证明BIK 1是一个双特异性激酶的第一次。使用定量质谱法（MS）明确评估BIK 1取代突变体的激酶活性。Thr-237、Thr-242和Tyr-250被发现在体外对BAK 1的自磷酸化和磷酸化的BIK 1活性影响最显著。构建BIK 1的结构模型以进一步阐明特定磷酸化残基的分子功能。我们还绘制了BIK 1对FLS 2磷酸化的新位点，这些位点与BAK 1不同。这些体外结果可以为更深入的体内研究提供新的假设，从而更深入地了解磷酸化如何促进BIK 1活化并介导下游信号传导特异性。
Arabidopsis BOTRYTIS-INDUCED KINASE1 (BIK1) is a receptor-like cytoplasmic kinase acting early in multiple signaling pathways important for plant growth and innate immunity. It is known to form a signaling complex with a cell-surface receptor FLS2 and a co-receptor kinase BAK1 to transduce signals upon perception of pathogen-associated molecular patterns (PAMPs). Although site-specific phosphorylation is speculated to mediate the activation and function of BIK1, few studies have been devoted to complete profiling of BIK1 phosphorylation residues. Here, we identified nineteen in vitro autophosphorylation sites of BIK1 including three phosphotyrosine sites, thereby proving BIK1 is a dual-specificity kinase for the first time. The kinase activity of BIK1 substitution mutants were explicitly assessed using quantitative mass spectrometry (MS). Thr-237, Thr-242 and Tyr-250 were found to most significantly affect BIK1 activity in autophosphorylation and phosphorylation of BAK1 in vitro. A structural model of BIK1 was built to further illustrate the molecular functions of specific phosphorylation residues. We also mapped new sites of FLS2 phosphorylation by BIK1, which are different from those by BAK1. These in vitro results could provide new hypotheses for more in-depth in vivo studies leading to deeper understanding of how phosphorylation contributes to BIK1 activation and mediates downstream signaling specificity.
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影响因子：
64.8
作者：
Chinchilla, Delphine;Zipfel, Cyril;Boller, Thomas
通讯作者：
Boller, Thomas
影响因子：
7
作者：
Boersema, Paul J.;Foong, Leong Yan;Heck, Albert J. R.
通讯作者：
Heck, Albert J. R.
DOI：
10.1073/pnas.0909705107
发表时间：
2010-01-05
影响因子：
11.1
作者：
Lu, Dongping;Wu, Shujing;He, Ping
通讯作者：
He, Ping
影响因子：
7.2
作者：
Kim, Dae Sung;Hwang, Byung Kook
通讯作者：
Hwang, Byung Kook
磷酸化对植物受体样激酶BAK1激活影响的结构基础
影响因子：
44.1
作者：
Jingwen Zhou;Zhiyong Wang;Wenqing Shui;Zhiyong Lou
通讯作者：
Zhiyong Lou