Synaptic activity bidirectionally regulates a novel sequence-specific S-Q phosphoproteome in neurons

J Neurochem. 2014 Mar;128(6):841-51. doi: 10.1111/jnc.12487. Epub 2013 Nov 13.

Abstract

Protein phosphorylation plays a critical role in neuronal transcription, translation, cell viability, and synaptic plasticity. In neurons, phospho-enzymes and specific substrates directly link glutamate release and post-synaptic depolarization to these cellular functions; however, many of these enzymes and their protein substrates remain uncharacterized or unidentified. In this article, we identify a novel, synaptically driven neuronal phosphoproteome characterized by a specific motif of serine/threonine-glutamine ([S/T]-Q, abbreviated as SQ). These SQ-containing substrates are predominantly localized to dendrites, synapses, the soma; and activation of this SQ phosphoproteome by bicuculline application is induced via calcium influx through L-type calcium channels. On the other hand, acute application of NMDA can inactivate this SQ phosphoproteome. We demonstrate that the SQ motif kinase Ataxia-telangiectasia mutated can also localize to dendrites and dendritic spines, in addition to other subcellular compartments, and is activated by bicuculline application. Pharmacology studies indicate that Ataxia-telangiectasia mutated and its sister kinase ataxia telangiectasia mutated and Rad3-related up-regulate these neuronal SQ substrates. Phosphoproteomics identified over 150 SQ-containing substrates whose phosphorylation is bidirectionally regulated by synaptic activity.

Keywords: L-type calcium channels; Phosphoproteomics; SQ motif; phosphorylation; synaptic activity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Action Potentials / drug effects
  • Action Potentials / physiology
  • Animals
  • Calcium / physiology
  • Calcium Channels, L-Type / physiology
  • Cerebral Cortex / cytology
  • Enzyme Inhibitors / pharmacology
  • Female
  • Male
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / physiology*
  • Organ Culture Techniques
  • Phosphoproteins / physiology*
  • Phosphorylation / physiology
  • Pregnancy
  • Primary Cell Culture
  • Proteome / physiology
  • Proteomics*
  • Rats
  • Sodium Channel Blockers / pharmacology
  • Synapses / drug effects
  • Synapses / physiology*
  • Tetrodotoxin / pharmacology

Substances

  • Calcium Channels, L-Type
  • Enzyme Inhibitors
  • Phosphoproteins
  • Proteome
  • Sodium Channel Blockers
  • Tetrodotoxin
  • Calcium