Protein S-Nitrosylation Controls Glycogen Synthase Kinase 3β Function Independent of Its Phosphorylation State.
Protein S-Nitrosylation Controls Glycogen Synthase Kinase 3β Function Independent of Its Phosphorylation State.
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DOI：
10.1161/circresaha.118.312789
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发表时间：
2018-05-25
影响因子：
20.1
通讯作者：
Van Eyk JE
中科院分区：
文献类型：
--
作者：
Wang SB;Venkatraman V;Crowgey EL;Liu T;Fu Z;Holewinski R;Ranek M;Kass DA;O'Rourke B;Van Eyk JE
糖原合成酶激酶3β（Glycogen synthase kinase 3β，GSK 3 β）是一种多功能的组成型激酶，参与多种细胞过程的调节。其功能调节的主要机制是通过丝氨酸-9残基的磷酸化依赖性抑制。新出现的证据表明，可能存在控制GSK 3 β某些功能的替代机制。在此，我们试图了解蛋白质S-亚硝基化（SNO）对GSK 3 β功能的作用。在体外研究了GSK 3定位的SNO依赖性调节及其磷酸化下游靶点的能力，还研究了心力衰竭发展过程中受磷酸化或SNO依赖性GSK 3调节和体内关键信号传导激酶的SNO修饰差异影响的蛋白质网络。我们发现，GSK 3 β在体外HEK 293细胞、H9 C2成肌细胞和原代新生大鼠心室肌细胞（NRVM）中以及在体内心力衰竭和心源性猝死动物模型的心脏中经历位点特异性SNO。GSK 3 β的S-亚硝基化显著抑制其激酶活性，不依赖于经典磷酸化抑制途径。GSK 3 β的S-亚硝基化促进其核转位并接近新的下游磷酸化底物，这些底物富含新的氨基酸共有序列基序。定量磷酸化蛋白质组学途径分析表明，细胞核中的GSK 3 β在细胞周期调控、RNA剪接和DNA损伤反应中起着重要作用。结果表明，SNO对GSK 3 β在细胞质和细胞核中的定位和活性具有不同的影响。GSK 3 β的SNO修饰发生在体内，并可能导致心力衰竭和心源性猝死的病理生物学。
Glycogen synthase kinase 3β (GSK3β) is a multifunctional and constitutively active kinase known to regulate a myriad of cellular processes. The primary mechanism to regulate its function is through phosphorylation-dependent inhibition at serine-9 residue. Emerging evidence indicates that there may be alternative mechanisms that control GSK3β for certain functions. Here we sought to understand the role of protein S-nitrosylation (SNO) on the function of GSK3β. SNO-dependent modulation of the localization of GSK3 and its ability to phosphorylate downstream targets was investigated in vitro and the network of proteins differentially impacted by phospho- or SNO-dependent GSK3 regulation and in vivo SNO modification of key signaling kinases during the development of heart failure was also studied. We found that GSK3β undergoes site-specific SNO both in vitro, in HEK293 cells, H9C2 myoblasts, and primary neonatal rat ventricular myocytes (NRVM), as well as in vivo, in hearts from an animal model of heart failure and sudden cardiac death. S-nitrosylation of GSK3β significantly inhibits its kinase activity independent of the canonical phospho-inhibition pathway. S-nitrosylation of GSK3β promotes its nuclear translocation and access to novel downstream phospho-substrates which are enriched for a novel amino acid consensus sequence motif. Quantitative phospho-proteomics pathway analysis reveals that nuclear GSK3β plays a central role in cell cycle control, RNA splicing and DNA damage response. The results indicate that SNO has a differential effect on the location and activity of GSK3β in the cytoplasm versus the nucleus. SNO modification of GSK3β occurs in vivo and could contribute to the pathobiology of heart failure and sudden cardiac death.