The Phosphatase PP2A Interacts With ArnA and ArnB to Regulate the Oligomeric State and the Stability of the ArnA/B Complex
The Phosphatase PP2A Interacts With ArnA and ArnB to Regulate the Oligomeric State and the Stability of the ArnA/B Complex
复制标题
磷酸酶PP 2A与ArnA和ArnB相互作用调节寡聚体状态和ArnA/B复合物的稳定性
DOI：
10.3389/fmicb.2020.01849
复制
发表时间：
2020-08-21
影响因子：
5.2
通讯作者：
Albers, Sonja-Verena
中科院分区：
文献类型：
--
作者：
Ye, Xing;Vogt, Marian Samuel;Albers, Sonja-Verena
在crenarchaeonSulfolobus acidocaldarius，古菌，IV型菌毛一样的运动结构，合成响应营养饥饿。古菌组分的合成受古菌调节网络（arn）控制。蛋白磷酸化在这个调控网络中起着重要的作用，因为编码蛋白激酶和磷酸酶PP 2A的几个基因的缺失影响细胞运动。古细菌调控网络中的几种蛋白质可以被磷酸化，然而，不同组分的磷酸化水平如何影响古细菌合成的细节仍然未知。鉴定与S.为了检测酸性热磷酸酶PTP和PP 2A的活性，进行了与质谱分析偶联的免疫共沉淀测定。在营养饥饿培养基中生长30分钟后，特别是保守的推定ATP/GTP结合蛋白（Saci_1281）、通用应激蛋白（Saci_0887）和古菌调节剂ArnA和ArnB被鉴定为PP 2A的高度丰富的相互作用蛋白。进一步研究了ArnA、ArnB和PP 2A之间的相互作用。以前的研究表明，含有Arn A的Forkhead相关结构域与含有Arn B的von Willebrand A型结构域相互作用，并且这两种蛋白质在体外都能被Arn C激酶磷酸化，纯化的蛋白质重组为Arn A/B异二聚体。在与ArnA的复合物中，ArnC激酶对ArnB的磷酸化被强烈刺激，并导致形成（ArnA/B）（2）和更高的寡聚复合物，而PP 2A的缔合和去磷酸化导致这些ArnA/B复合物的解离。
In the crenarchaeonSulfolobus acidocaldarius, the archaellum, a type-IV pilus like motility structure, is synthesized in response to nutrient starvation. Synthesis of components of the archaellum is controlled by thearchaellumregulatorynetwork (arn). Protein phosphorylation plays an important role in this regulatory network since the deletion of several genes encoding protein kinases and the phosphatase PP2A affected cell motility. Several proteins in the archaellum regulatory network can be phosphorylated, however, details of how phosphorylation levels of different components affect archaellum synthesis are still unknown. To identify proteins interacting with theS. acidocaldariusphosphatases PTP and PP2A, co-immunoprecipitation assays coupled to mass spectrometry analysis were performed. Thirty minutes after growth in nutrient starvation medium, especially a conserved putative ATP/GTP binding protein (Saci_1281), a universal stress protein (Saci_0887) and the archaellum regulators ArnA and ArnB were identified as highly abundant interaction proteins of PP2A. The interaction between ArnA, ArnB, and PP2A was further studied. Previous studies showed that the Forkhead-associated domain containing ArnA interacts with von Willebrand type A domain containing ArnB, and that both proteins could be phosphorylated by the kinase ArnCin vitro.The ArnA/B heterodimer was reconstituted from the purified proteins. In complex with ArnA, phosphorylation of ArnB by the ArnC kinase was strongly stimulated and resulted in formation of (ArnA/B)(2)and higher oligomeric complexes, while association and dephosphorylation by PP2A resulted in dissociation of these ArnA/B complexes.