The neuropeptide Y/peptide YY (PYY) Y1 receptor subtype mediates proliferative responses. This report identifies effector molecules which mediate mitogen-activated protein kinase (MAPK) phosphorylation by Y1 receptor activation in transfected CHO cells. Pertussis toxin pretreatment abolishes this effect, indicating involvement of G_ior G_oproteins. Inhibition of protein kinase C (PKC) also blocks PYY-induced MAPK phosphorylation. Additionally in this cell model PYY causes an increase in GTP binding to Ras protein, and cotransfection of dominant negative constructs for Ras and Raf blocks PYY effects on MAPK. These data suggest a novel mechanism for Y1 receptor coupling to MAPK, which is at once pertussis toxin-sensitive as well as PKC- and Ras-dependent.