TY - JOUR
T1 - Protein S-Nitrosylation Controls Glycogen Synthase Kinase 3β Function Independent of Its Phosphorylation State
AU - Wang, Sheng Bing
AU - Venkatraman, Vidya
AU - Crowgey, Erin L.
AU - Liu, Ting
AU - Fu, Zongming
AU - Holewinski, Ronald
AU - Ranek, Mark
AU - Kass, David A.
AU - O'Rourke, Brian
AU - Van Eyk, Jennifer E.
N1 - Funding Information:
This study was funded by National Institutes of Health (NIH)-National Heart, Lung, and Blood Institute (NHLBI) Program Project Grants on Pathology of Cardiac Dysynchrony and Resynchronization, P01HL77189-01, to J.E.Van Eyk, B. O’Rourke, and D.A. Kass. NIH-NHLBI contract on The Johns Hopkins Innovation Proteomics Center in Heart Failure, NHLBI-HV-10-05 (2), to J.E.Van Eyk, B. O’Rourke, and D.A. Kass. We also acknowledge the Barbara Streisand Women’s Heart Center (to J.E.Van Eyk), The Smidt Heart Institute at Cedars-Sinai Medical Center (to J.E.Van Eyk), and The Erika Glazer Endowed Chair in Women’s Heart Health (to J.E.Van Eyk) to support this study.
Publisher Copyright:
© 2018 American Heart Association, Inc.
PY - 2018/5/25
Y1 - 2018/5/25
N2 - Rationale: GSK-3β (glycogen synthase kinase 3β) 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 GSK-3β for certain functions. Objectives: Here, we sought to understand the role of protein S-nitrosylation (SNO) on the function of GSK-3β. SNO-dependent modulation of the localization of GSK-3β and its ability to phosphorylate downstream targets was investigated in vitro, and the network of proteins differentially impacted by phospho-or SNO-dependent GSK-3β regulation and in vivo SNO modification of key signaling kinases during the development of heart failure was also studied. Methods and Results: We found that GSK-3β undergoes site-specific SNO both in vitro, in HEK293 cells, H9C2 myoblasts, and primary neonatal rat ventricular myocytes, as well as in vivo, in hearts from an animal model of heart failure and sudden cardiac death. S-nitrosylation of GSK-3β significantly inhibits its kinase activity independent of the canonical phospho-inhibition pathway. S-nitrosylation of GSK-3β promotes its nuclear translocation and access to novel downstream phosphosubstrates which are enriched for a novel amino acid consensus sequence motif. Quantitative phosphoproteomics pathway analysis reveals that nuclear GSK-3β plays a central role in cell cycle control, RNA splicing, and DNA damage response. Conclusions: The results indicate that SNO has a differential effect on the location and activity of GSK-3β in the cytoplasm versus the nucleus. SNO modification of GSK-3β occurs in vivo and could contribute to the pathobiology of heart failure and sudden cardiac death.
AB - Rationale: GSK-3β (glycogen synthase kinase 3β) 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 GSK-3β for certain functions. Objectives: Here, we sought to understand the role of protein S-nitrosylation (SNO) on the function of GSK-3β. SNO-dependent modulation of the localization of GSK-3β and its ability to phosphorylate downstream targets was investigated in vitro, and the network of proteins differentially impacted by phospho-or SNO-dependent GSK-3β regulation and in vivo SNO modification of key signaling kinases during the development of heart failure was also studied. Methods and Results: We found that GSK-3β undergoes site-specific SNO both in vitro, in HEK293 cells, H9C2 myoblasts, and primary neonatal rat ventricular myocytes, as well as in vivo, in hearts from an animal model of heart failure and sudden cardiac death. S-nitrosylation of GSK-3β significantly inhibits its kinase activity independent of the canonical phospho-inhibition pathway. S-nitrosylation of GSK-3β promotes its nuclear translocation and access to novel downstream phosphosubstrates which are enriched for a novel amino acid consensus sequence motif. Quantitative phosphoproteomics pathway analysis reveals that nuclear GSK-3β plays a central role in cell cycle control, RNA splicing, and DNA damage response. Conclusions: The results indicate that SNO has a differential effect on the location and activity of GSK-3β in the cytoplasm versus the nucleus. SNO modification of GSK-3β occurs in vivo and could contribute to the pathobiology of heart failure and sudden cardiac death.
KW - S-nitrosylation
KW - glycogen synthase kinase 3 beta
KW - kinase-substrates interactome
KW - nuclear translocation
KW - redox regulation
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U2 - 10.1161/CIRCRESAHA.118.312789
DO - 10.1161/CIRCRESAHA.118.312789
M3 - Article
C2 - 29563102
AN - SCOPUS:85052398423
SN - 0009-7330
VL - 122
SP - 1517
EP - 1531
JO - Circulation research
JF - Circulation research
IS - 11
ER -