application) The long-term goal of this research has been to elucidate the function of specific protein kinase C (PKC) isozymes in the control of vascular smooth muscle (VSM) cell contractility and growth. An understanding of these signaling pathways could provide a basis for explaining or treating the pathology underlying pervasive vascular diseases including hypertension, atherosclerosis, and restenosis. The mitogen-activated protein kinase (MAPK) signaling pathway is essential for controlling cell growth and gene transcription and has also been implicated in the control of VSM cell motility and contractility. Studies indicate that a pool of VSM cell PKC-delta, a Ca2+-independent isozyme of PKC, and Raf-1 (an upstream activator of the MAPK) co-immunoprecipitate, suggesting a physical interaction.
One specific aim the proposed research is to test the hypothesis that the pool of interacting PKC-delta and Raf-1 is coupled functionally (i.e. PKC-delta activation leads to stimulation of Raf serine/threonine kinase activity) and determine to what degree this contributes to MAPK activation in response to tyrosine kinase receptor-coupled growth factors and G-protein receptor coupled ligands. Tyrosine phosphorylated focal adhesion kinase (FAK) and paxillin, also co-immunoprecipitate with PKC-delta from VSM cell lysates. This has led to the hypothesis that PKC-delta and Raf physically interact with focal adhesion complexes, which are structures required for cell adhesion, migration, and force transmission.
The second aim of the research is to determine: A) whether integrin-dependent MAPK signaling stimulated by cell adhesion or stretch is mediated specifically by the pool of interacting PKC-delta/Raf; B) whether the interaction of PKC-delta and Raf with focal adhesion complex proteins is modulated by integrin-dependent VSM cell adhesion and/or stretch of adherent cells; and C) whether PKC-delta regulates integrin-dependent VSM cell adhesion or migration.
The final aim i s to determine if Src- or a related tyrosine-kinase phosphorylates PKC-delta thereby regulating PKC-delta /Raf/focal adhesion complex interactions and the integrin-dependent signaling pathway. This research will lead to a better understanding of the mechanisms by which PKC regulates focal adhesion assembly/disassembly and provide insight into the integrin-dependent signaling pathways which control VSM cell migration and patterns of gene expression.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL040992-11
Application #
2702182
Study Section
Experimental Cardiovascular Sciences Study Section (ECS)
Project Start
1988-07-01
Project End
2001-08-31
Budget Start
1998-09-01
Budget End
1999-08-31
Support Year
11
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Albany Medical College
Department
Physiology
Type
Schools of Medicine
DUNS #
City
Albany
State
NY
Country
United States
Zip Code
12208
Ginnan, Roman; Guikema, Benjamin J; Halligan, Katharine E et al. (2008) Regulation of smooth muscle by inducible nitric oxide synthase and NADPH oxidase in vascular proliferative diseases. Free Radic Biol Med 44:1232-45
Ginnan, Roman; Guikema, Benjamin J; Singer, Harold A et al. (2006) PKC-delta mediates activation of ERK1/2 and induction of iNOS by IL-1beta in vascular smooth muscle cells. Am J Physiol Cell Physiol 290:C1583-91
Ginnan, Roman; Singer, Harold A (2005) PKC-delta-dependent pathways contribute to PDGF-stimulated ERK1/2 activation in vascular smooth muscle. Am J Physiol Cell Physiol 288:C1193-201
Ginnan, Roman; Pfleiderer, Paul J; Pumiglia, Kevin et al. (2004) PKC-delta and CaMKII-delta 2 mediate ATP-dependent activation of ERK1/2 in vascular smooth muscle. Am J Physiol Cell Physiol 286:C1281-9
Ginnan, Roman; Singer, Harold A (2002) CaM kinase II-dependent activation of tyrosine kinases and ERK1/2 in vascular smooth muscle. Am J Physiol Cell Physiol 282:C754-61
Rokolya, A; Walsh, M P; Singer, H A et al. (1998) Protein kinase C--catalyzed calponin phosphorylation in swine carotid arterial homogenate. J Cell Physiol 176:545-52
Bhat, G J; Abraham, S T; Singer, H A et al. (1997) Alpha-thrombin stimulates sis-inducing factor-A DNA binding activity in rat aortic smooth muscle cells. Hypertension 29:356-60
Busuttil, S J; Morehouse, D L; Youkey, J R et al. (1996) Antisense suppresssion of protein kinase C-alpha and -delta in vascular smooth muscle. J Surg Res 63:137-42
Schorb, W; Conrad, K M; Singer, H A et al. (1995) Angiotensin II is a potent stimulator of MAP-kinase activity in neonatal rat cardiac fibroblasts. J Mol Cell Cardiol 27:1151-60
Booz, G W; Taher, M M; Baker, K M et al. (1994) Angiotensin II induces phosphatidic acid formation in neonatal rat cardiac fibroblasts: evaluation of the roles of phospholipases C and D. Mol Cell Biochem 141:135-43

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