APRO4 negatively regulates Src tyrosine kinase activity in PC12 cells.

The Src nonreceptor tyrosine kinase plays an important role in multiple signalling pathways that regulate several cellular functions including proliferation, differentiation and transformation. The activity of Src is tightly regulated in vivo and can be modulated by interactions of its SH2 and SH3 domains with high-affinity ligands. APRO4 (anti-proliferative 4) belongs to a new antiproliferative gene family involved in the negative control of the cell cycle. This report shows that APRO4 associates with Src via its C-terminal proline-rich domain, and downregulates Src kinase activity. Moreover, overexpression of APRO4 leads to inhibition of neurite outgrowth and Ras/MAP kinase signalling in PC12 cells. Furthermore, the kinetics of endogenous Src inactivation correlates with an increase in endogenous APRO4 co-immunoprecipitation in FGF-stimulated PC12 cells. Finally, downregulation of endogenous APRO4 by expression of antisense RNA induces the activation of Src and spontaneous formation of neurites in PC12 cells. Therefore, by controlling the basal threshold of Src activity, APRO4 constitutes an important negative regulatory mechanism for Src-mediated signalling.