beta-Adrenergic Signaling Inhibits G(q)-Dependent Protein Kinase D Activation by Preventing Protein Kinase D Translocation

C. Blake Nichols, Chia-Wei Chang, Maura Ferrero, Brittani M. Wood, Matthew L. Stein, Amanda J. Ferguson, Derrick Ha, Robert R. Rigor, Sven Bossuyt, Julie Bossuyt*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

8 Citations (Scopus)

Abstract

Rationale: Both -adrenergic receptor (-AR) and G(q)-coupled receptor (G(q)R) agonist-driven signaling play key roles in the events, leading up to and during cardiac dysfunction. How these stimuli interact at the level of protein kinase D (PKD), a nodal point in cardiac hypertrophic signaling, remains unclear.

Objective: To assess the spatiotemporal dynamics of PKD activation in response to -AR signaling alone and on coactivation with G(q)R-agonists. This will test our hypothesis that compartmentalized PKD signaling reconciles disparate findings of PKA facilitation and inhibition of PKD activation.

Methods and Results: We report on the spatial and temporal profiles of PKD activation using green fluorescent protein-tagged PKD (wildtype or mutant S427E) and targeted fluorescence resonance energy transfer-based biosensors (D-kinase activity reporters) in adult cardiomyocytes. We find that -AR/PKA signaling drives local nuclear activation of PKD, without preceding sarcolemmal translocation. We also discover pronounced interference of -AR/cAMP/PKA signaling on G(q)R-induced translocation and activation of PKD throughout the cardiomyocyte. We attribute these effects to direct, PKA-dependent phosphorylation of PKD-S427. We also show that phosphomimetic substitution of S427 likewise impedes G(q)R-induced PKD translocation and activation. In neonatal myocytes, S427E inhibits G(q)R-evoked cell growth and expression of hypertrophic markers. Finally, we show altered S427 phosphorylation in transverse aortic constriction-induced hypertrophy.

Conclusions: -AR signaling triggers local nuclear signaling and inhibits G(q)R-mediated PKD activation by preventing its intracellular translocation. PKA-dependent phosphorylation of PKD-S427 fine-tunes the PKD responsiveness to G(q)R-agonists, serving as a key integration point for -adrenergic and G(q)-coupled stimuli.

Original languageEnglish
Pages (from-to)1398-1409
Number of pages12
JournalCIRCULATION RESEARCH
Volume114
Issue number9
DOIs
Publication statusPublished - 25 Apr 2014
MoE publication typeA1 Journal article-refereed

Keywords

  • cyclic AMP-dependent protein kinases
  • GTP-binding proteins
  • myocytes
  • cardiac
  • protein kinase D
  • receptors
  • adrenergic
  • CARDIAC TROPONIN-I
  • PHOSPHOLIPASE-C
  • CARDIOVASCULAR-SYSTEM
  • HISTONE DEACETYLASE-5
  • ADULT CARDIOMYOCYTES
  • VENTRICULAR MYOCYTES
  • PHOSPHORYLATION
  • STIMULATION
  • HYPERTROPHY
  • RECEPTOR

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