Angiotensin II Stimulates p21-Activated Kinase in Vascular Smooth Muscle Cells : Role in Activation of JNK

Department of Medicine, University of Washington, Seattle 98195, USA.
Circulation Research (Impact Factor: 11.02). 07/1998; 82(12):1272-8. DOI: 10.1161/01.RES.82.12.1272
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Angiotensin II (Ang II) has been previously shown to stimulate the extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK) mitogen-activated protein (MAP) kinase family members. Little is known regarding the upstream signaling molecules involved in Ang II-mediated JNK activation. Ang II has been shown to activate the Janus kinase/signal transducer(s) and activator(s) of transcription (JAK/STAT) pathway, suggesting similarities to cytokine signaling. In response to cytokines such as interleukin-1 and tumor necrosis factor-alpha, the p21-activated kinase (PAK) has been identified as an upstream component in JNK activation. Therefore, we hypothesized that PAK may be involved in JNK activation by Ang II in vascular smooth muscle cells (VSMCs). AlphaPAK activity was measured by myelin basic protein phosphorylation in rat aortic VSMCs. In response to Ang II, alphaPAK was rapidly stimulated within 1 minute, with a peak (5-fold increase) at 30 minutes. AlphaPAK stimulation preceded activation of JNK in VSMCs. Ang II-mediated activation of both alphaPAK and JNK was Ca2+ dependent and inhibited by downregulation of phorbol ester-sensitive protein kinase C isoforms (by pretreatment with phorbol 12,13-dibutyrate) but not by pretreatment with GF109203X. Activation of both PAK and JNK was partially inhibited by tyrosine kinase inhibitors but not by specific Src inhibitors, suggesting regulation by a tyrosine kinase other than c-Src. Finally, introduction of dominant negative PAK markedly reduced the JNK activation by Ang II in both Chinese hamster ovary and COS cells stably expressing the Ang II type 1 receptor (AT1R). Our data provide evidence for alphaPAK as an upstream mediator of JNK in Ang II signaling and extend the role of Ang II as a proinflammatory mediator for VSMCs.

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Available from: Udo Schmitz, Aug 03, 2015
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    • "A large number of studies of the functions of Ang II in vitro and in vivo have indicated that Ang II activates multiple intracellular signaling cascades, such as those mediated by mitogen-activated protein kinase (MAPK), and regulates various transcription factors, and causes cell growth in vascular smooth muscle cells (VSMCs)1, 2, 3. A growing body of evidence shows that Ang II type 1 (AT-1) receptor but not AT-2 is involved in the signal transduction underlying Ang II-induced VSMC growth1, such as activation of JNK4 and p385. However, little is known about the role of these signal cascades in Ang II-induced proliferative response. "
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    • "Studies in a variety of Ang II target cell types have shown that the Ang II activation of different pathways is time dependent. For example, activation of the G-protein-dependent pathway and generation of IP 3 occurs in seconds, while MAPK and JAK/STAT activation occurs in minutes to hours after initial AT1R activation (Ishida et al., 1995; Schmitz et al., 1998). While many of the Ang II signaling cascades have been defined, an understanding of the downstream changes in gene expression has moved ahead very slowly, often through the definition of one gene-target at a time. "
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    • "JNK kinase assay was performed as described previously [18]. Briefly, approximately 250 Ag of cell lysate protein was incubated with 3 Ag GST-c-Jun (1-169) coupled to glutathione agarose at 4 -C for 4 h under constant rotation. "
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