ARRDC3 tyrosine phosphorylation functions as a switch to control c-Src versus WWP2 interactions and distinct scaffolding functions

ABSTRACT: Mammalian -arrestins are members of the same arrestin family as the ubiquitously expressed and extensively studied -arrestins. Arrestins share common structural elements including the conserved N- and C-arrestin-fold domains, polar core, finger loop, and C-terminal tail, all of which mediate protein-protein interactions. In -arrestins, these domains enable the control of G protein-coupled receptors (GPCR) signaling and scaffolding interactions with various signaling proteins including c-Src. By contrast, the repertoire of -arrestin scaffolding partners and regulatory mechanisms that control their interactions are not well understood. -arrestins differ considerably from -arrestins in the C-terminal region; -arrestins contain clathrin adaptor -adaptin binding sites whereas -arrestins harbor PPxY motifs, demonstrated to interact with WW domains of E3 ubiquitin ligases such as Itch and WWP2. Here we report the identification of a novel phosphorylation site, tyrosine (Y) 394, embedded in the C-terminal PPxY motif of -arrestin ARRDC3. The Y394 site functions as a phospho-regulatory switch to enable distinct ARRDC3 binding partners and scaffolding functions. We demonstrate that ARRDC3 Y394 phosphorylation promotes interaction with c-Src via its SH2 domain, whereas the non-phosphorylated form preferentially binds to WWP2. Our results further show that ARRDC3 Y394 phosphorylation and c-Src SH2 domain-dependent interaction enables regulation of c-Src activity, whereas ARRDC3 Y394 phosphorylation disrupts WWP2 interaction and perturbs ARRDC3-dependent lysosomal trafficking of the GPCR, protease-activated receptor-1. Together these findings indicate that ARRDC3 Y394 functions as a phospho-regulatory switch to enable selective binding to different partners that impact distinct scaffolding functions.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hek-293t Cell

SUBMITTER: Majid Ghassemian  

LAB HEAD: JoAnn Trejo

PROVIDER: PXD060782 | Pride | 2025-07-14

REPOSITORIES: Pride