G-protein-coupled receptor kinase interactor-1 serine 419 accelerates premature synapse formation in cortical neurons by interacting with Ca(2+)/calmodulin-dependent protein kinase IIβ

Brain Res Bull. 2013 Jun:95:70-7. doi: 10.1016/j.brainresbull.2013.01.006. Epub 2013 Jan 24.

Abstract

In the present study, we investigated the mechanisms of brain derived neurotrophic factor (BDNF) in regulating cortical neuron premature synapse formation. KN-93, a specific inhibitor of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), and G-protein-coupled receptor kinase interactor-1 (G1T1) siRNA were utilized, and the premature synapse formation of cortical neurons was detected under BDNF stimulation. Plasmids HA-GIT1, HA-GIT1 (ΔSLD), HA-GIT1 (S419A) and Flag-CaMKIIβ were constructed. The interaction between GIT1 and CaMKIIβ, and their influence on the premature synapse formation of BDNF-stimulated cortical neurons were examined. BDNF-stimulated cortical neurons were associated with increased premature synapse formation, the enhancement of phosphorylation for CaMKIIβ, and the combination of GIT1 and p-CaMKII(thr286). G1T1 siRNA and KN-93 inhibited premature synapse formation in cortical neurons. The interaction between GIT1 and CaMKIIβ required SLD domain and serine 419 in GIT1. BDNF-induced CaMKIIβ phosphorylation and premature synapse formation were suppressed in GIT1 (S419A) transfected cortical neurons. By interacting with CaMKIIβ, G1T1 (S419) were shown to participate in BDNF-induced premature synapse formation within cortical neurons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzylamines
  • Brain-Derived Neurotrophic Factor / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / metabolism*
  • Calmodulin / metabolism*
  • Cell Cycle Proteins / metabolism*
  • Cells, Cultured
  • G-Protein-Coupled Receptor Kinases / antagonists & inhibitors*
  • Neurons / metabolism
  • Phosphoproteins / metabolism*
  • Rats
  • Serine / metabolism
  • Signal Transduction / physiology
  • Sulfonamides
  • Synapses / metabolism*

Substances

  • Benzylamines
  • Brain-Derived Neurotrophic Factor
  • Calmodulin
  • Cell Cycle Proteins
  • Git1 protein, rat
  • Phosphoproteins
  • Sulfonamides
  • KN 93
  • Serine
  • G-Protein-Coupled Receptor Kinases
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2