JAK2 regulates Nav1.6 channel function via FGF14Y158 phosphorylation

Paul A. Wadsworth, Aditya K. Singh, Nghi Nguyen, Nolan M. Dvorak, Cynthia M. Tapia, William K. Russell, Clifford Stephan, Fernanda Laezza

Research output: Contribution to journalArticle

Abstract

Background: Protein interactions between voltage-gated sodium (Nav) channels and accessory proteins play an essential role in neuronal firing and plasticity. However, a surprisingly limited number of kinases have been identified as regulators of these molecular complexes. We hypothesized that numerous as-of-yet unidentified kinases indirectly regulate the Nav channel via modulation of the intracellular fibroblast growth factor 14 (FGF14), an accessory protein with numerous unexplored phosphomotifs and required for channel function in neurons. Methods: Here we present results from an in-cell high-throughput screening (HTS) against the FGF14: Nav1.6 complex using >3000 diverse compounds targeting an extensive range of signaling pathways. Regulation by top kinase targets was then explored using in vitro phosphorylation, biophysics, mass-spectrometry and patch-clamp electrophysiology. Results: Compounds targeting Janus kinase 2 (JAK2) were over-represented among HTS hits. Phosphomotif scans supported by mass spectrometry revealed FGF14Y158, a site previously shown to mediate both FGF14 homodimerization and interactions with Nav1.6, as a JAK2 phosphorylation site. Following inhibition of JAK2, FGF14 homodimerization increased in a manner directly inverse to FGF14:Nav1.6 complex formation, but not in the presence of the FGF14Y158A mutant. Patch-clamp electrophysiology revealed that through Y158, JAK2 controls FGF14-dependent modulation of Nav1.6 channels. In hippocampal CA1 pyramidal neurons, the JAK2 inhibitor Fedratinib reduced firing by a mechanism that is dependent upon expression of FGF14. Conclusions: These studies point toward a novel mechanism by which levels of JAK2 in neurons could directly influence firing and plasticity by controlling the FGF14 dimerization equilibrium, and thereby the availability of monomeric species for interaction with Nav1.6.

Original languageEnglish (US)
Article number118786
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1867
Issue number10
DOIs
StatePublished - Oct 2020

Keywords

  • FGF14
  • JAK2
  • Signaling
  • Sodium channels
  • Tyrosine kinases

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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