THE NITRIC OXIDE SYNTHASE ADAPTOR PROTEIN (NOS1AP) ASSOCIATES WITH SCRIBBLE AND REGULATES DENDRITIC SPINE DEVELOPMENT

Abstract

In a targeted proteomic screen to identify polarity protein complexes, a number of Scribble (Scrib) -associating proteins were identified; of particular interest was the Nitric Oxide Synthase 1 Adaptor Protein (NOS1AP). NOS1AP contains an N-terminal phosphotyrosine binding (PTB) domain and a C-terminal PSD-95/Dlg homology/ZO-1 (PDZ) binding motif that associates with neuronal NOS (NOS1). We show that the PTB domain of NOS1AP associates with the fourth PDZ domain of Scrib. We identified NOS1AP binding partners including three key regulators of dendritic spine formation, beta-Pix, Git1, and PAK, which require Scrib to associate with NOS1AP. Overexpression of NOS1AP in cultured hippocampal neurons increases dendritic protrusions, a process dependent on the PTB domain. The increase in dendritic protrusions can be blocked by the co-expression of a dominant negative Rac construct. NOS1AP, and the PTB domain of NOS1AP influence Rac activity. Together these data suggest that Scrib and NOS1AP function as important scaffolding proteins in the mammalian synapse and that NOS1AP functions in the dendritic spine by influencing Rho GTPase activity.