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.
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.
Department of Spine Surgery, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China.
Zitao
Z
Zhang
Wei
W
Zhou
Jin
J
Fan
Yongxin
Y
Ren
Guoyong
G
Yin
D000818:Animals; D001596:Benzylamines; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D054768:G-Protein-Coupled Receptor Kinases / Q000037:antagonists & inhibitors*; D009474:Neurons / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D051381:Rats; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D013449:Sulfonamides; D013569:Synapses / Q000378:metabolism*; D000096926:Benzenesulfonamides
D001596:Benzylamines; D019208:Brain-Derived Neurotrophic Factor; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D002147:Calmodulin; D018797:Cell Cycle Proteins; D054768:G-Protein-Coupled Receptor Kinases; D010750:Phosphoproteins; D012694:Serine; D013449:Sulfonamides; C485648:Git1 protein, rat; C072105:KN 93; D000096926:Benzenesulfonamides