Protein S-Nitrosylation Controls Glycogen Synthase Kinase 3β Function Independent of Its Phosphorylation State

Shengbing Wang, Vidya Venkatraman, Erin L. Crowgey, Ting Liu, Zongming Fu, Ronald Holewinski, Mark Ranek, David A Kass, Brian O'Rourke, Jennifer E. Van Eyk

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1517-1531
Number of pages15
JournalCirculation Research
Volume122
Issue number11
DOIs
Publication statusPublished - May 25 2018
Externally publishedYes

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Keywords

  • glycogen synthase kinase 3 beta
  • kinase-substrates interactome
  • nuclear translocation
  • redox regulation
  • S-nitrosylation

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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