MPK12 in stomatal CO2 signaling: function beyond its kinase activity
MPK12 in stomatal CO2 signaling: function beyond its kinase activity
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MPK12 在气孔 CO2 信号传导中的作用：其功能超出其激酶活性
DOI：
10.1111/nph.18913
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发表时间：
2023
期刊：
影响因子：
9.4
通讯作者：
Garcia‐Sosa, Alfonso T.
中科院分区：
文献类型：
--
作者：
Yeh, Chung‐Yueh;Wang, Yuh‐Shuh;Takahashi, Yohei;Kuusk, Katarina;Paul, Karnelia;Arjus, Triinu;Yadlos, Oleksii;Schroeder, Julian I.;Ilves, Ivar;Garcia‐Sosa, Alfonso T.
蛋白质磷酸化是调节气孔开闭的一个重要分子开关。先前的研究将MAP激酶12和Raf样激酶HT 1之间的相互作用定义为CO2浓度变化引起的气孔运动的必要步骤。然而，MPK 12激酶活性是否是CO2诱导的气孔反应调控所必需的，还有待深入研究。我们应用遗传学、生物化学和结构模拟方法，研究了MPK 12在保卫细胞CO2信号传导中的非催化作用，该作用依赖于HT 1的变构抑制。通过分析在MPK 12缺失的植物系中表达MPK 12的各种激酶死亡和组成型活性版本的植物系的气体交换，我们证实了CO2依赖性气孔反应依赖于MPK 12结合HT 1的能力，而不是其激酶活性。我们还证明了纯化的MPK 12和HT 1蛋白在CO2/HCO 3 −存在下形成异源二聚体，并提出了解释MPK 12：HT 1相互作用界面的结构模型。这些数据补充了MPK 12激酶活性无关的与HT 1的相互作用作为分子开关的模型，通过该开关，保卫细胞感知大气CO2浓度的变化。
Protein phosphorylation is a major molecular switch involved in the regulation of stomatal opening and closure. Previous research defined interaction between MAP kinase 12 and Raf‐like kinase HT1 as a required step for stomatal movements caused by changes in CO2concentration. However, whether MPK12 kinase activity is required for regulation of CO2‐induced stomatal responses warrants in‐depth investigation.We apply genetic, biochemical, and structural modeling approaches to examining the noncatalytic role of MPK12 in guard cell CO2signaling that relies on allosteric inhibition of HT1.We show that CO2/HCO3−‐enhanced MPK12 interaction with HT1 is independent of its kinase activity. By analyzing gas exchange of plant lines expressing various kinase‐dead and constitutively active versions of MPK12 in a plant line whereMPK12is deleted, we confirmed that CO2‐dependent stomatal responses rely on MPK12's ability to bind to HT1, but not its kinase activity. We also demonstrate that purified MPK12 and HT1 proteins form a heterodimer in the presence of CO2/HCO3−and present structural modeling that explains the MPK12:HT1 interaction interface.These data add to the model that MPK12 kinase‐activity‐independent interaction with HT1 functions as a molecular switch by which guard cells sense changes in atmospheric CO2concentration.
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