PKG1α Cysteine-42 Redox State Controls mTORC1 Activation in Pathological Cardiac Hypertrophy

Christian U. Oeing, Taishi Nakamura, Shi Pan, Sumita Mishra, Brittany L. Dunkerly-Eyring, Kristen M. Kokkonen-Simon, Brian L. Lin, Anna Chen, Guangshuo Zhu, Djahida Bedja, Dong Ik Lee, David A. Kass, Mark Ranek

Research output: Contribution to journalArticle

Abstract

Rationale: Stimulated PKG1α (protein kinase G-1α) phosphorylates TSC2 (tuberous sclerosis complex 2) at serine 1365, potently suppressing mTORC1 (mechanistic [mammalian] target of rapamycin complex 1) activation by neurohormonal and hemodynamic stress. This reduces pathological hypertrophy and dysfunction and increases autophagy. PKG1α oxidation at cysteine-42 is also induced by these stressors, which blunts its cardioprotective effects. Objective: We tested the dependence of mTORC1 activation on PKG1α C42 oxidation and its capacity to suppress such activation by soluble GC-1 (guanylyl cyclase 1) activation. Methods and Results: Cardiomyocytes expressing wild-type (WT) PKG1α (PKG1αWT) or cysteine-42 to serine mutation redox-dead (PKG1αCS/CS) were exposed to ET-1 (endothelin 1). Cells expressing PKG1αWTexhibited substantial mTORC1 activation (p70 S6K [p70 S6 kinase], 4EBP1 [elF4E binding protein-1], and Ulk1 [Unc-51-like kinase 1] phosphorylation), reduced autophagy/autophagic flux, and abnormal protein aggregation; all were markedly reversed by PKG1αCS/CSexpression. Mice with global knock-in of PKG1αCS/CSsubjected to pressure overload (PO) also displayed markedly reduced mTORC1 activation, protein aggregation, hypertrophy, and ventricular dysfunction versus PO in PKG1αWTmice. Cardioprotection against PO was equalized between groups by co-treatment with the mTORC1 inhibitor everolimus. TSC2-S1365 phosphorylation increased in PKG1αCS/CSmore than PKG1αWTmyocardium following PO. TSC2S1365A/S1365A(TSC2 S1365 phospho-null, created by a serine to alanine mutation) knock-in mice lack TSC2 phosphorylation by PKG1α, and when genetically crossed with PKG1αCS/CSmice, protection against PO-induced mTORC1 activation, cardiodepression, and mortality in PKG1αCS/CSmice was lost. Direct stimulation of GC-1 (BAY-602770) offset disparate mTORC1 activation between PKG1αWTand PKG1αCS/CSafter PO and blocked ET-1 stimulated mTORC1 in TSC2S1365A-expressing myocytes. Conclusions: Oxidation of PKG1α at C42 reduces its phosphorylation of TSC2, resulting in amplified PO-stimulated mTORC1 activity and associated hypertrophy, dysfunction, and depressed autophagy. This is ameliorated by direct GC-1 stimulation.

Original languageEnglish (US)
Pages (from-to)522-533
Number of pages12
JournalCirculation research
DOIs
StateAccepted/In press - 2020

Keywords

  • autophagy
  • heart failure
  • hypertrophy
  • mice
  • phosphorylation

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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    Oeing, C. U., Nakamura, T., Pan, S., Mishra, S., Dunkerly-Eyring, B. L., Kokkonen-Simon, K. M., Lin, B. L., Chen, A., Zhu, G., Bedja, D., Lee, D. I., Kass, D. A., & Ranek, M. (Accepted/In press). PKG1α Cysteine-42 Redox State Controls mTORC1 Activation in Pathological Cardiac Hypertrophy. Circulation research, 522-533. https://doi.org/10.1161/CIRCRESAHA.119.315714