Calcium/calmodulin-dependent protein kinase II phosphorylates and regulates the Drosophila eag potassium channel.
Modulation of neuronal excitability is believed to be an important mechanism of plasticity in the nervous system. Calcium/calmodulin-dependent protein kinase II (CaMKII) has been postulated to regulate the ether à go-go (eag) potassium channel in Drosophila. Inhibition of CaMKII and mutation of the eag gene both cause hyperexcitability at the larval neuromuscular junction (NMJ) and memory formation defects in the adult. In this study, we identify a single site, threonine 787, as the major CaMKII phosphorylation site in Eag. This site can be phosphorylated by CaMKII both in a heterologous cell system and in vivo at the larval NMJ. Expression of Eag in Xenopus oocytes was used to assess the function of phosphorylation. Injection of either a specific CaMKII inhibitor peptide or lavendustin C, another CaMKII inhibitor, reduced Eag current amplitude acutely. Mutation of threonine 787 to alanine also reduced amplitude. Moreover, both CaMKII inhibition and the alanine mutation accelerated inactivation. The reduction in current amplitudes and the accelerated inactivation of dephosphorylated Eag channels would result in decreased outward potassium currents and hyperexcitability at presynaptic terminals and, thus, are consistent with the NMJ phenotype observed when CaMKII is inhibited. These results show that Eag is a substrate of CaMKII and suggest that direct modulation of potassium channels may be an important function of this kinase.
Wang
Zheng
Z
Department of Biology and Volen Center for Complex Systems, Brandeis University, Waltham, Massachusetts 02454-9110, USA.
Wilson
Gisela F
GF
Griffith
Leslie C
LC
D000818:Animals; D001665:Binding Sites; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000201:enzymology / Q000502:physiology*; D051638:Ether-A-Go-Go Potassium Channels; D005786:Gene Expression Regulation; D007814:Larva; D009469:Neuromuscular Junction / Q000502:physiology; D010766:Phosphorylation; D015221:Potassium Channels / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
D029721:Drosophila Proteins; D051638:Ether-A-Go-Go Potassium Channels; D015221:Potassium Channels; D011994:Recombinant Proteins; C496883:eag protein, Drosophila; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases