Arai Ken, Maruyama Yoshiko, Nishida Motohiro, Tanabe Shihori, Takagahara Shuichi, Kozasa Tohru, Mori Yasuo, Nagao Taku, Kurose Hitoshi
Laboratory of Pharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.
Mol Pharmacol. 2003 Mar;63(3):478-88. doi: 10.1124/mol.63.3.478.
In the present study, we examined the roles of G(12), G(13), G(q), and G(i) in endothelin-1-induced hypertrophic responses. Endothelin-1 stimulation activated extracellular signal-regulated kinase (ERK) and c-Jun NH(2)-terminal kinase (JNK) in cultured rat neonatal myocytes. The activation of JNK, but not ERK, was inhibited by the expression of carboxyl terminal regions of G alpha(12) and G alpha(13). JNK activation was also inhibited by expression of the G alpha(12)/G alpha(13)-specific inhibitor regulator of G protein signaling (RGS) domain of p115RhoGEF and the G alpha(q)-specific inhibitor RGS domain of the G protein-coupled receptor kinase 2 (GRK2-RGS). JNK activation was not, however, inhibited by expression of the carboxyl terminal region of G protein-coupled receptor kinase 2 (GRK2-ct), which is a G beta gamma-sequestering polypeptide. Additionally, JNK activation but not ERK activation was inhibited by the expression of C3 exoenzyme that inactivates small GTPase Rho. These results suggest that JNK activation by G alpha(12), G alpha(13), and G alpha(q) is involved in Rho. On the other hand, ERK activation was inhibited by pertussis toxin treatment, the receptor-G(i) uncoupler, and GRK2-ct. Thus, ERK was activated by G alpha(i)- and G beta gamma-dependent pathways. These results clearly demonstrate that differential pathways activate JNK and ERK.
在本研究中,我们检测了G(12)、G(13)、G(q)和G(i)在内皮素-1诱导的肥厚反应中的作用。内皮素-1刺激可激活培养的大鼠新生心肌细胞中的细胞外信号调节激酶(ERK)和c-Jun氨基末端激酶(JNK)。Gα(12)和Gα(13)羧基末端区域的表达可抑制JNK的激活,而不是ERK的激活。p115RhoGEF的Gα(12)/Gα(13)特异性抑制剂G蛋白信号调节因子(RGS)结构域和G蛋白偶联受体激酶2(GRK2-RGS)的Gα(q)特异性抑制剂RGS结构域的表达也可抑制JNK的激活。然而,G蛋白偶联受体激酶2(GRK2-ct)羧基末端区域的表达并不能抑制JNK的激活,GRK2-ct是一种Gβγ隔离多肽。此外,使小GTP酶Rho失活的C3外切酶的表达可抑制JNK的激活,但不能抑制ERK的激活。这些结果表明,Gα(12)、Gα(13)和Gα(q)对JNK的激活与Rho有关。另一方面,百日咳毒素处理、受体-G(i)解偶联剂和GRK2-ct可抑制ERK的激活。因此,ERK是由Gα(i)和Gβγ依赖性途径激活的。这些结果清楚地表明,JNK和ERK是通过不同途径激活的。