Angiotensin II-induced transactivation of epidermal growth factor receptor regulates fibronectin and transforming growth factor-beta synthesis via transcriptional and posttranscriptional mechanisms.

The signaling cascade elicited by angiotensin II (Ang II) resembles that characteristic of a growth factor, and recent evidence indicates transactivation of epidermal growth factor receptor (EGF-R) by G protein-coupled receptors. Here, we report the involvement of EGF-R in Ang II-induced synthesis of fibronectin and transforming growth factor-beta (TGF-beta) in cardiac fibroblasts. Ang II stimulated fibronectin mRNA levels dose dependently, with a maximal increase (approximately 5-fold) observed after 12 hours of incubation. Fibronectin synthesis induced by Ang II or calcium ionophore was completely abolished by tyrosine kinase inhibitors and intracellular Ca2+ chelating agents. Ang II-induced fibronectin mRNA was not affected by protein kinase C inhibitors or protein kinase C depletion, whereas specific inhibition of EGF-R function by a dominant negative EGF-R mutant and tyrphostin AG1478 abolished induction of fibronectin mRNA. We isolated the rat fibronectin gene, including the 5'-flanking region, and found that the activator protein-1 (AP-1) binding site present in the promoter region was responsible for the Ang II responsiveness of this gene. A gel retardation assay revealed the binding of nuclear protein to the AP-1 site, which was supershifted with anti-c-fos and anti-c-jun but not anti-activating transcription factor (ATF)-2 antibodies. Conditioned medium from Ang II-treated cells contained TGF-beta bioactivity, and addition of neutralizing TGF-beta antibody modestly (46%) inhibited induction of fibronectin. Ang II-induced synthesis of TGF-beta was also abolished by inhibition of EGF-R function. The effect of TGF-beta was exerted by stabilizing fibronectin mRNA without affecting the promoter activity and required de novo protein synthesis. We concluded that Ang II-induced expression of fibronectin and TGF-beta is mediated by downstream signaling of EGF-R transactivated by Ca2+-dependent tyrosine kinase and that Ang II-induced fibronectin mRNA expression is regulated by 2 different mechanisms, which are transcriptional control by binding of the c-fos/c-jun complex to the AP-1 site and posttranscriptional control by mRNA stabilization due to autocrine or paracrine effects of TGF-beta. Thus, this study suggests that the action of Ang II on extracellular matrix formation should be interpreted in association with the EGF-R signaling cascade.

[1]  Y. Mori,et al.  Role of calcium-sensitive tyrosine kinase Pyk2/CAKbeta/RAFTK in angiotensin II induced Ras/ERK signaling. , 1998, Hypertension.

[2]  M. Shibuya,et al.  Angiotensin II type 1 receptor-induced extracellular signal-regulated protein kinase activation is mediated by Ca2+/calmodulin-dependent transactivation of epidermal growth factor receptor. , 1998, Circulation research.

[3]  H. Kawakatsu,et al.  Calcium-dependent Epidermal Growth Factor Receptor Transactivation Mediates the Angiotensin II-induced Mitogen-activated Protein Kinase Activation in Vascular Smooth Muscle Cells* , 1998, The Journal of Biological Chemistry.

[4]  H. Matsubara,et al.  Molecular insights into angiotensin II type 1 and type 2 receptors: expression, signaling and physiological function and clinical application of its antagonists. , 1998, Endocrine journal.

[5]  Tsonwin Hai,et al.  Epidermal Growth Factor Induction of the c-jun Promoter by a Rac Pathway , 1998, Molecular and Cellular Biology.

[6]  Y. Mori,et al.  Angiotensin type 2 receptors are reexpressed by cardiac fibroblasts from failing myopathic hamster hearts and inhibit cell growth and fibrillar collagen metabolism. , 1997, Circulation.

[7]  A. Ullrich,et al.  Signal characteristics of G protein‐transactivated EGF receptor , 1997, The EMBO journal.

[8]  A. Ullrich,et al.  Critical Role of Calcium- dependent Epidermal Growth Factor Receptor Transactivation in PC12 Cell Membrane Depolarization and Bradykinin Signaling* , 1997, The Journal of Biological Chemistry.

[9]  J. Schlessinger,et al.  Phosphatidylinositol 3-kinase mediates epidermal growth factor-induced activation of the c-Jun N-terminal kinase signaling pathway , 1997, Molecular and cellular biology.

[10]  E. Peralta,et al.  The m1 muscarinic acetylcholine receptor transactivates the EGF receptor to modulate ion channel activity , 1997, The EMBO journal.

[11]  M. Marrero,et al.  Angiotensin II Stimulates Tyrosine Phosphorylation and Activation of Insulin Receptor Substrate 1 and Protein-tyrosine Phosphatase 1D in Vascular Smooth Muscle Cells* , 1997, The Journal of Biological Chemistry.

[12]  B. Berk,et al.  Angiotensin II signal transduction in vascular smooth muscle: role of tyrosine kinases. , 1997, Circulation research.

[13]  R. Lefkowitz,et al.  Gβγ Subunits Mediate Src-dependent Phosphorylation of the Epidermal Growth Factor Receptor , 1997, The Journal of Biological Chemistry.

[14]  R. Lefkowitz,et al.  Gbetagamma subunits mediate Src-dependent phosphorylation of the epidermal growth factor receptor. A scaffold for G protein-coupled receptor-mediated Ras activation. , 1997, The Journal of biological chemistry.

[15]  Y. Zou,et al.  Angiotensin II stimulates c-Jun NH2-terminal kinase in cultured cardiac myocytes of neonatal rats. , 1997, Circulation research.

[16]  S. Kudoh,et al.  Protein Kinase C, but Not Tyrosine Kinases or Ras, Plays a Critical Role in Angiotensin II-induced Activation of Raf-1 Kinase and Extracellular Signal-regulated Protein Kinases in Cardiac Myocytes* , 1996, The Journal of Biological Chemistry.

[17]  Y. Mori,et al.  Down-regulation by cAMP of angiotensin II type 2 receptor gene expression in PC12 cells. , 1996, Hypertension research : official journal of the Japanese Society of Hypertension.

[18]  IsseiKomuro,et al.  Mechanical Stretch Induces Enhanced Expression of Angiotensin II Receptor Subtypes in Neonatal Rat Cardiac Myocytes , 1996 .

[19]  Y. Mori,et al.  Mechanical stretch induces enhanced expression of angiotensin II receptor subtypes in neonatal rat cardiac myocytes. , 1996, Circulation research.

[20]  M. Marrero,et al.  The Importance of Tyrosine Phosphorylation in Angiotensin II Signaling. , 1996, Trends in cardiovascular medicine.

[21]  S. Eguchi,et al.  Identification of an Essential Signaling Cascade for Mitogen-activated Protein Kinase Activation by Angiotensin II in Cultured Rat Vascular Smooth Muscle Cells , 1996, The Journal of Biological Chemistry.

[22]  J. Avruch,et al.  Stress-activated protein kinases in cardiovascular disease. , 1996, Circulation research.

[23]  M. Karin.  The regulation of AP-1 activity by mitogen-activated protein kinases. , 1996, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[24]  A. Ullrich,et al.  Role of transactivation of the EGF receptor in signalling by G-protein-coupled receptors , 1996, Nature.

[25]  M. Greenberg,et al.  Stimulation of growth factor receptor signal transduction by activation of voltage-sensitive calcium channels. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[26]  J. Sadoshima,et al.  The heterotrimeric G q protein‐coupled angiotensin II receptor activates p21 ras via the tyrosine kinase‐Shc‐Grb2‐Sos pathway in cardiac myocytes. , 1996, The EMBO journal.

[27]  M. Karin,et al.  Induction of c‐fos expression through JNK‐mediated TCF/Elk‐1 phosphorylation. , 1995, The EMBO journal.

[28]  Y. Mori,et al.  Identification of a Negative Cis-regulatory Element and Trans-acting Protein That Inhibit Transcription of the Angiotensin II Type 1a Receptor Gene (*) , 1995, The Journal of Biological Chemistry.

[29]  M. Karin.  The Regulation of AP-1 Activity by Mitogen-activated Protein Kinases (*) , 1995, The Journal of Biological Chemistry.

[30]  R. Treisman,et al.  The Rho family GTPases RhoA, Racl , and CDC42Hsregulate transcriptional activation by SRF , 1995, Cell.

[31]  C. Benjamin,et al.  Convergence of Angiotensin II and Platelet-derived Growth Factor Receptor Signaling Cascades in Vascular Smooth Muscle Cells (*) , 1995, The Journal of Biological Chemistry.

[32]  B. Berk,et al.  Direct stimulation of Jak/STAT pathway by the angiotensin II AT1 receptor , 1995, Nature.

[33]  S. Fisher,et al.  Norepinephrine and ANG II stimulate secretion of TGF-beta by neonatal rat cardiac fibroblasts in vitro. , 1995, The American journal of physiology.

[34]  A. Levitzki,et al.  Tyrosine kinase inhibition: an approach to drug development. , 1995, Science.

[35]  C. Slaughter,et al.  ERK phosphorylation potentiates Elk‐1‐mediated ternary complex formation and transactivation. , 1995, The EMBO journal.

[36]  J. Sadoshima,et al.  Angiotensin II and Other Hypertrophic Stimuli Mediated by G Protein–Coupled Receptors Activate Tyrosine Kinase, Mitogen-Activated Protein Kinase, and 90-kD S6 Kinase in Cardiac Myocytes The Critical Role of Ca2+-Dependent Signaling , 1995 .

[37]  J. Sadoshima,et al.  Angiotensin II and other hypertrophic stimuli mediated by G protein-coupled receptors activate tyrosine kinase, mitogen-activated protein kinase, and 90-kD S6 kinase in cardiac myocytes. The critical role of Ca(2+)-dependent signaling. , 1995, Circulation research.

[38]  S. Keyse,et al.  Tyrosine kinase inhibition: an approach to drug development. , 1995, Human & experimental toxicology.

[39]  H. Matsubara,et al.  Regulation of gene transcription of angiotensin II receptor subtypes in myocardial infarction. , 1995, The Journal of clinical investigation.

[40]  G. J. Bhat,et al.  Angiotensin II stimulates sis-inducing factor-like DNA binding activity. Evidence that the AT1A receptor activates transcription factor-Stat91 and/or a related protein. , 1994, The Journal of biological chemistry.

[41]  H. Singer,et al.  Involvement of protein kianse C and Ca2+ in angiotensin II-induced mitogenesis of cardiac fibroblasts. , 1994, The American journal of physiology.

[42]  W. Schorb,et al.  Angiotensin II-induced protein tyrosine phosphorylation in neonatal rat cardiac fibroblasts. , 1994, The Journal of biological chemistry.

[43]  S. Kagami,et al.  Angiotensin II stimulates extracellular matrix protein synthesis through induction of transforming growth factor-beta expression in rat glomerular mesangial cells. , 1994, The Journal of clinical investigation.

[44]  P. Erne,et al.  Characterization of angiotensin II receptors in cultured adult rat cardiac fibroblasts. Coupling to signaling systems and gene expression. , 1994, The Journal of clinical investigation.

[45]  H. Matsubara,et al.  Differential gene expression and regulation of angiotensin II receptor subtypes in rat cardiac fibroblasts and cardiomyocytes in culture. , 1994, The Journal of clinical investigation.

[46]  R. Treisman.  Ternary complex factors: growth factor regulated transcriptional activators. , 1994, Current opinion in genetics & development.

[47]  T. Inagami,et al.  Angiotensin II Receptor: Molecular Cloning, Functions, and Regulation , 1994 .

[48]  H. Matsubara,et al.  Regulatory elements that mediate expression of the gene for the angiotensin II type 1a receptor for the rat. , 1993, The Journal of biological chemistry.

[49]  Noel N Kim,et al.  Identification of Functional Angiotensin II Receptors on Rat Cardiac Fibroblasts , 1993, Circulation.

[50]  H. Matsubara,et al.  Rat angiotensin II (type 1A) receptor mRNA regulation and subtype expression in myocardial growth and hypertrophy. , 1993, Circulation research.

[51]  G. Booz,et al.  Angiotensin II is mitogenic in neonatal rat cardiac fibroblasts. , 1993, Circulation research.

[52]  C. Molloy,et al.  Angiotensin II stimulation of rapid protein tyrosine phosphorylation and protein kinase activation in rat aortic smooth muscle cells. , 1993, The Journal of biological chemistry.

[53]  K. Shii,et al.  Angiotensin II stimulates two myelin basic protein/microtubule-associated protein 2 kinases in cultured vascular smooth muscle cells. , 1992, Circulation research.

[54]  G. Gibbons,et al.  Vascular smooth muscle cell hypertrophy vs. hyperplasia. Autocrine transforming growth factor-beta 1 expression determines growth response to angiotensin II. , 1992, The Journal of clinical investigation.

[55]  P. Brecher,et al.  Fibronectin expression in the normal and hypertrophic rat heart. , 1992, The Journal of clinical investigation.

[56]  D. Gardner,et al.  Divergent regulation of the human atrial natriuretic peptide gene by c-jun and c-fos , 1992, Molecular and cellular biology.

[57]  M. Eghbali,et al.  Differential effects of transforming growth factor-beta 1 and phorbol myristate acetate on cardiac fibroblasts. Regulation of fibrillar collagen mRNAs and expression of early transcription factors. , 1991, Circulation research.

[58]  K. Weber,et al.  Pathological Hypertrophy and Cardiac Interstitium: Fibrosis and Renin‐Angiotensin‐Aldosterone System , 1991, Circulation.

[59]  K. Watanabe,et al.  Specific inhibitors of tyrosine-specific protein kinases: properties of 4-hydroxycinnamamide derivatives in vitro. , 1989, Cancer research.

[60]  H. Moses,et al.  Regulation of mRNAs for type-1 plasminogen activator inhibitor, fibronectin, and type I procollagen by transforming growth factor-beta. Divergent responses in lung fibroblasts and carcinoma cells. , 1988, The Journal of biological chemistry.

[61]  M. Sporn,et al.  A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-β , 1988, Cell.

[62]  D. Brown,et al.  An improved method of purification of transforming growth factor, type beta from platelets. , 1988, Analytical biochemistry.

[63]  R. Hynes,et al.  Organization of the fibronectin gene provides evidence for exon shuffling during evolution. , 1987, The EMBO journal.

[64]  M. Karin,et al.  Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor , 1987, Cell.