The Src family tyrosine kinase is involved in Rho-dependent activation of c-Jun N-terminal kinase by Gα12

G alpha(12), a member of alpha subunit of heterotrimeric G protein G(12) subfamily, has been shown to stimulate c-Jun N-terminal kinase (JNK) activity through the low molecular weight GTP-binding proteins Ras, Rac, and Cdc42. In this study using the transient expression of a constitutively activated mutant of G alpha(12) (G alpha(12)Q229L) in human embryonic kidney (HEK) 293 cells, we found that Rho and Src family kinase are also involved in the Gu,,induced activation of JNK. The activation of JNK by G alpha(12)Q229L was inhibited by dominant-negative RhoA(T19N), and botulinum C3 exoenzyme which specifically inactivates Rho. In addition, the expression of activated RhoA(G14V) elevated JNK activity in HEK 293 cells. The G alpha(12)Q229L-stimulated activation of JNK was blocked by a specific inhibitor of protein tyrosine kinases (PP2), and C-terminal Src kinase (Csk). Moreover, we observed that G alpha(12)Q229L stimulated Src family kinase activity and v-Src induced JNK activation. Interestingly, the v-Src-induced activation of JNK was inhibited by dominant-negative RhoA(T19N). In contrast, Csk did not inhibit the JNK activation by activated RhoA(G14V). These results suggest that Rho and Src family kinase are required for the G alpha(12)-induced JNK activation, and that Src family kinase acts upstream of Rho activation in the JNK pathway.