title: Calcium/calmodulin-dependent protein kinase II phosphorylation drives synapse-associated protein 97 into spines.
abstract: Synapse-associated protein 97 (SAP97) has been involved in the correct delivery and clustering of glutamate ionotropic receptors to the postsynaptic compartment. Here we demonstrate that synaptic trafficking of SAP97 itself was modulated by calcium/calmodulin-dependent protein kinase II (CaMKII) in cultured hippocampal neurons. CaMKII activation led to increased targeting of SAP97 into dendritic spines, whereas CaMKII inhibition was responsible for SAP97 high colocalization in the cell soma with the endoplasmic reticulum protein disulfide-isomerase. No effect was detected for other members of the membrane-associated guanylate kinase protein family, such as SAP102 and PSD-95. Transfection of activated alphaCaMKII T286D dramatically increased concentration of both endogenous and transfected SAP97 at postsynaptic terminals. In vitro CaMKII phosphorylation of the SAP97 N-terminal fusion protein and metabolic labeling of transfected COS7 cells indicated SAP97-Ser-39 as a CaMKII phosphosite in the SAP97 protein sequence. Moreover, transfection in hippocampal neurons of SAP97 mutants that blocked or mimicked Ser-39 phosphorylation had effects similar to those observed upon inhibiting or constitutively activating CaMKII. Further, CaMKII-dependent SAP97-Ser-39 phosphorylation determined a redistribution of the glutamate receptor subunit (GluR1) of the AMPA receptor. In conclusion, our data show that CaMKII-dependent SAP97-Ser-39 phosphorylation regulates the association of SAP97 with the postsynaptic complex, thus providing a fine molecular mechanism responsible for the synaptic delivery of SAP97 interacting proteins, i.e. ionotropic glutamate receptor subunits.
authors.affiliation: Center of Excellence on Neurodegenerative Diseases and Department of Pharmacological Sciences, University of Milano, via Balzaretti 9, 20133 Milano, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Mauceri
authors.forename: Flaminio
authors.initials: F
authors.lastname: Cattabeni
authors.forename: Monica
authors.initials: M
authors.lastname: Di Luca
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Gardoni
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000502:physiology*; D002478:Cells, Cultured; D006624:Hippocampus / Q000378:metabolism*; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins / Q000502:physiology*; D009474:Neurons / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D013569:Synapses / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C501119:Dlg1 protein, rat; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases