Role of Calcium-Sensitive Tyrosine Kinase Pyk2/CAK beta/RAFTK in Angiotensin II-Induced Ras/ERK Signaling

• Published in: Hypertension (Dallas, Tex. 1979) Tex. 1979), 1998-10, Vol.32 (4), p.668-675
• Main Authors: Murasawa, Satoshi, Mori, Yasukiyo, Nozawa, Yoshihisa, Masaki, Hiroya, Maruyama, Katsuya, Tsutsumi, Yoshiaki, Moriguchi, Yasutaka, Shibasaki, Yasunobu, Tanaka, Yoko, Iwasaka, Toshiji, Inada, Mitsuo, Matsubara, Hiroaki
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Heart; Vertebrates: cardiovascular system
• ISSN: 0194-911X; 1524-4563
• DOI: 10.1161/01.HYP.32.4.668

In cardiac fibroblasts, angiotensin II (Ang II) induced a rapid increase in extracellular signal-regulated kinase (ERK) activity in a pertussis toxin-insensitive manner. This ERK activation was abolished by the Gq-associated phospholipase C inhibitor U73122 but was insensitive to protein kinase C (PKC) inhibitors or PKC downregulation by phorbol ester. Intracellular Ca chelation by BAPTA-AM or TMB-8 abolished Ang II-induced ERK activation, whereas treatment with EGTA or nifedipine did not affect it. Ca ionophore A23187 also induced a rapid increase in ERK activity to an extent similar to that of Ang II stimulation. Calmodulin inhibitors (W7 and calmidazolium) and tyrosine kinase inhibitors (genistein and ST638) completely blocked ERK activation by Ang II and A23187. Both Ang II and A23187 caused a rapid increase in the binding of GTP to p21, which was nearly abolished by genistein and calmidazolium. Transfection with the dominant negative mutant of Ras and the Ras inhibitor manumycin completely inhibited Ang II-induced ERK activation. It was also found for the first time that cardiac fibroblasts abundantly expressed Ca tyrosine kinase Pyk2/CAK beta/RAFTK and that Ang II markedly induced its activation in a Ca manner. Overexpression of the dominant negative mutant of Pyk2 significantly attenuated Ang II- or A23187-induced ERK activities (36% and 38% inhibition compared with that in mock-transfected cells, respectively) and ERK tyrosine phosphorylation levels, as well as an increase in the binding of GTP to p21. These findings demonstrate that in cardiac fibroblasts, Ang II-induced Ras/ERK activation is dominantly regulated by Gq-coupled Ca signaling and that Pyk2 plays an important role in the signal transmission for efficient activation of the Ang II-induced Ras/ERK pathway. (Hypertension. 1998;32:668-675.)