Cardiac-specific overexpression of AT1 receptor mutant lacking Gαq/Gαi coupling causes hypertrophy and bradycardia in transgenic mice

Ang II type 1 (AT1) receptors activate both conventional heterotrimeric G protein-dependent and unconventional G protein-independent mechanisms. We investigated how these different mechanisms activated by AT1 receptors affect growth and death of cardiac myocytes in vivo. Transgenic mice with cardiac-specific overexpression of WT AT1 receptor (AT1-WT; Tg-WT mice) or an AT1 receptor second intracellular loop mutant (AT1-i2m; Tg-i2m mice) selectively activating G(alpha)q/G(alpha)i-independent mechanisms were studied. Tg-i2m mice developed more severe cardiac hypertrophy and bradycardia coupled with lower cardiac function than Tg-WT mice. In contrast, Tg-WT mice exhibited more severe fibrosis and apoptosis than Tg-i2m mice. Chronic Ang II infusion induced greater cardiac hypertrophy in Tg-i2m compared with Tg-WT mice whereas acute Ang II administration caused an increase in heart rate in Tg-WT but not in Tg-i2m mice. Membrane translocation of PKCepsilon, cytoplasmic translocation of G(alpha)q, and nuclear localization of phospho-ERKs were observed only in Tg-WT mice while activation of Src and cytoplasmic accumulation of phospho-ERKs were greater in Tg-i2m mice, consistent with the notion that G(alpha)q/G(alpha)i-independent mechanisms are activated in Tg-i2m mice. Cultured myocytes expressing AT1-i2m exhibited a left and upward shift of the Ang II dose-response curve of hypertrophy compared with those expressing AT1-WT. Thus, the AT1 receptor mediates downstream signaling mechanisms through G(alpha)q/G(alpha)i-dependent and -independent mechanisms, which induce hypertrophy with a distinct phenotype.

[1]  S. Vatner,et al.  Silent Information Regulator 2&agr;, a Longevity Factor and Class III Histone Deacetylase, Is an Essential Endogenous Apoptosis Inhibitor in Cardiac Myocytes , 2004, Circulation research.

[2]  W. D. De Mello,et al.  Intracellular and Extracellular Angiotensin II Enhance the L-Type Calcium Current in the Failing Heart , 2004, Hypertension.

[3]  J. L. Hansen,et al.  Oligomerization of Wild Type and Nonfunctional Mutant Angiotensin II Type I Receptors Inhibits Gαq Protein Signaling but Not ERK Activation* , 2004, Journal of Biological Chemistry.

[4]  S. Kudoh,et al.  Mechanical stress activates angiotensin II type 1 receptor without the involvement of angiotensin II , 2004, Nature Cell Biology.

[5]  M. Hori,et al.  Ca(2+)-sensitive tyrosine kinase Pyk2/CAK beta-dependent signaling is essential for G-protein-coupled receptor agonist-induced hypertrophy. , 2004, Journal of molecular and cellular cardiology.

[6]  G. Breithardt,et al.  Gene dose-dependent atrial arrhythmias, heart block, and brady-cardiomyopathy in mice overexpressing A(3) adenosine receptors. , 2004, Cardiovascular research.

[7]  W. Giles,et al.  Nkx2-5 Pathways and Congenital Heart Disease Loss of Ventricular Myocyte Lineage Specification Leads to Progressive Cardiomyopathy and Complete Heart Block , 2004, Cell.

[8]  H. Katus,et al.  Phosphorylation of Eukaryotic Translation Initiation Factor 2Bε by Glycogen Synthase Kinase-3β Regulates β-Adrenergic Cardiac Myocyte Hypertrophy , 2004, Circulation research.

[9]  R. Lefkowitz,et al.  Reciprocal Regulation of Angiotensin Receptor-activated Extracellular Signal-regulated Kinases by β-Arrestins 1 and 2* , 2004, Journal of Biological Chemistry.

[10]  J. Ramos,et al.  RSK2 Activity Is Regulated by Its Interaction with PEA-15* , 2003, Journal of Biological Chemistry.

[11]  Sábata S Constancio,et al.  Focal Adhesion Kinase Is Activated and Mediates the Early Hypertrophic Response to Stretch in Cardiac Myocytes , 2003, Circulation research.

[12]  S. Fluharty,et al.  Structural determinants for the activation mechanism of the angiotensin II type 1 receptor differ for phosphoinositide hydrolysis and mitogen-activated protein kinase pathways. , 2003, Biochemical pharmacology.

[13]  S. Vatner,et al.  Activation of Mst1 causes dilated cardiomyopathy by stimulating apoptosis without compensatory ventricular myocyte hypertrophy. , 2003, The Journal of clinical investigation.

[14]  J. Sadoshima,et al.  Phosphorylation of Tyrosine 319 of the Angiotensin II Type 1 Receptor Mediates Angiotensin II-induced Trans-activation of the Epidermal Growth Factor Receptor* , 2003, The Journal of Biological Chemistry.

[15]  Susan R. George,et al.  G-Protein-coupled receptor oligomerization and its potential for drug discovery , 2002, Nature Reviews Drug Discovery.

[16]  G. Dorn,et al.  Mitochondrial death protein Nix is induced in cardiac hypertrophy and triggers apoptotic cardiomyopathy , 2002, Nature Medicine.

[17]  Zhao Zhang,et al.  Functional Roles of Ca(v)1.3 (alpha(1D)) calcium channel in sinoatrial nodes: insight gained using gene-targeted null mutant mice. , 2002, Circulation research.

[18]  M. Lew,et al.  Side-chain substitutions within angiotensin II reveal different requirements for signaling, internalization, and phosphorylation of type 1A angiotensin receptors. , 2002, Molecular pharmacology.

[19]  R. Lefkowitz,et al.  β-Arrestin Scaffolding of the ERK Cascade Enhances Cytosolic ERK Activity but Inhibits ERK-mediated Transcription following Angiotensin AT1a Receptor Stimulation* , 2002, The Journal of Biological Chemistry.

[20]  R. Neubig,et al.  AT1 Receptor Mutant Lacking Heterotrimeric G Protein Coupling Activates the Src-Ras-ERK Pathway without Nuclear Translocation of ERKs* , 2002, The Journal of Biological Chemistry.

[21]  K. Tamura,et al.  The angiotensin II type I receptor-associated protein, ATRAP, is a transmembrane protein and a modulator of angiotensin II signaling. , 2002, Molecular biology of the cell.

[22]  G. Milligan,et al.  Protein-protein interactions at G-protein-coupled receptors. , 2001, Trends in pharmacological sciences.

[23]  P. Hamet,et al.  The angiotensin II type 1 receptor and receptor-associated proteins , 2001, Cell Research.

[24]  R. Neubig,et al.  ANG II type 1 receptor downregulation does not require receptor endocytosis or G protein coupling. , 2001, American journal of physiology. Cell physiology.

[25]  E. Formstecher,et al.  PEA-15 mediates cytoplasmic sequestration of ERK MAP kinase. , 2001, Developmental cell.

[26]  J. Gutkind,et al.  G-protein-coupled receptors and signaling networks: emerging paradigms. , 2001, Trends in pharmacological sciences.

[27]  K. Bernstein,et al.  Tyrosine Kinase Activation by the Angiotensin II Receptor in the Absence of Calcium Signaling* , 2001, The Journal of Biological Chemistry.

[28]  H. Duff,et al.  Complete heart block and sudden death in mice overexpressing calreticulin. , 2001, The Journal of clinical investigation.

[29]  H. Cingolani,et al.  Angiotensin II stimulates cardiac L-type Ca(2+) current by a Ca(2+)- and protein kinase C-dependent mechanism. , 2001, American journal of physiology. Heart and circulatory physiology.

[30]  John W. Adams,et al.  G-proteins in growth and apoptosis: lessons from the heart , 2001, Oncogene.

[31]  T. Inagami,et al.  The Renin-Angiotensin System in the Twenty-first Century , 2001, Blood pressure.

[32]  R. Kitsis,et al.  The MEK1–ERK1/2 signaling pathway promotes compensated cardiac hypertrophy in transgenic mice , 2000, The EMBO journal.

[33]  H. Lother,et al.  AT1-receptor heterodimers show enhanced G-protein activation and altered receptor sequestration , 2000, Nature.

[34]  J. Engel,et al.  Congenital Deafness and Sinoatrial Node Dysfunction in Mice Lacking Class D L-Type Ca2+ Channels , 2000, Cell.

[35]  P. Kang,et al.  The conserved phosphoinositide 3‐kinase pathway determines heart size in mice , 2000, The EMBO journal.

[36]  N. Dali-Youcef,et al.  Overexpression of angiotensin II type I receptor in cardiomyocytes induces cardiac hypertrophy and remodeling. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[37]  G. Dorn,et al.  Cardiac-specific overexpression of Galphaq alters excitation-contraction coupling in isolated cardiac myocytes. , 1999, Journal of molecular and cellular cardiology.

[38]  J Ross,et al.  Cardiac-specific overexpression of RhoA results in sinus and atrioventricular nodal dysfunction and contractile failure. , 1999, The Journal of clinical investigation.

[39]  R. Lefkowitz,et al.  Heptahelical Receptor Signaling: Beyond the G Protein Paradigm , 1999, The Journal of cell biology.

[40]  P. Ping,et al.  PKC-dependent activation of p44/p42 MAPKs during myocardial ischemia-reperfusion in conscious rabbits. , 1999, American journal of physiology. Heart and circulatory physiology.

[41]  J. Bockaert,et al.  Molecular tinkering of G protein‐coupled receptors: an evolutionary success , 1999, The EMBO journal.

[42]  D. Roden,et al.  Replacement by homologous recombination of the minK gene with lacZ reveals restriction of minK expression to the mouse cardiac conduction system. , 1999, Circulation research.

[43]  B. Conklin,et al.  Conditional expression and signaling of a specifically designed Gi-coupled receptor in transgenic mice , 1999, Nature Biotechnology.

[44]  M. Marrero,et al.  Regulation of angiotensin II-induced JAK2 tyrosine phosphorylation: roles of SHP-1 and SHP-2. , 1998, American journal of physiology. Cell physiology.

[45]  J. Girault,et al.  Endothelin Induces a Calcium‐Dependent Phosphorylation of PEA‐15 in Intact Astrocytes: Identification of Ser104 and Ser116 Phosphorylated, Respectively, by Protein Kinase C and Calcium/Calmodulin Kinase II In Vitro , 1998, Journal of neurochemistry.

[46]  P. Sugden,et al.  Oncogenic src, raf, and rasStimulate a Hypertrophic Pattern of Gene Expression and Increase Cell Size in Neonatal Rat Ventricular Myocytes* , 1998, The Journal of Biological Chemistry.

[47]  J. Sadoshima.  Versatility of the angiotensin II type 1 receptor. , 1998, Circulation research.

[48]  G. Dorn,et al.  Decompensation of Pressure-Overload Hypertrophy in Gαq-Overexpressing Mice , 1998 .

[49]  M. Marrero,et al.  Angiotensin II-induced Association of Phospholipase Cγ1 with the G-protein-coupled AT1 Receptor* , 1998, The Journal of Biological Chemistry.

[50]  M. Marrero,et al.  Dependence on the Motif YIPP for the Physical Association of Jak2 Kinase with the Intracellular Carboxyl Tail of the Angiotensin II AT1 Receptor* , 1997, The Journal of Biological Chemistry.

[51]  G. Barsh,et al.  Overexpression of angiotensin AT1 receptor transgene in the mouse myocardium produces a lethal phenotype associated with myocyte hyperplasia and heart block. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[52]  E. Kranias,et al.  Phospholamban deficiency alters inactivation kinetics of L-type Ca2+ channels in mouse ventricular myocytes. , 1997, The American journal of physiology.

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

[54]  T. Sakmar,et al.  Characterization of Rhodopsin Mutants That Bind Transducin but Fail to Induce GTP Nucleotide Uptake , 1995, The Journal of Biological Chemistry.

[55]  P. Mannon,et al.  Regulation of blood pressure by the type 1A angiotensin II receptor gene. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[56]  T. Katada,et al.  Mapping of G protein coupling sites of the angiotensin II type 1 receptor. , 1995, Hypertension.

[57]  W. Mello.  Is an intracellular renin-angiotensin system involved in control of cell communication in heart? , 1994 .

[58]  J. Downward,et al.  A complex of Grb2 adaptor protein, Sos exchange factor, and a 36-kDa membrane-bound tyrosine phosphoprotein is implicated in ras activation in T cells. , 1994, The Journal of biological chemistry.

[59]  J. Sadoshima,et al.  Autocrine release of angiotensin II mediates stretch-induced hypertrophy of cardiac myocytes in vitro , 1993, Cell.

[60]  S. Chaki,et al.  Domains for G-protein coupling in angiotensin II receptor type I: studies by site-directed mutagenesis. , 1992, Biochemical and biophysical research communications.

[61]  M. Schambelan,et al.  Characterization of angiotensin II receptor subtypes in rat heart. , 1992, Circulation research.

[62]  K. Rothblum,et al.  Intracardiac detection of angiotensinogen and renin: a localized renin-angiotensin system in neonatal rat heart. , 1992, The American journal of physiology.

[63]  A. Allen,et al.  In vitro autoradiographic localization of binding to angiotensin receptors in the rat heart. , 1990, International journal of cardiology.

[64]  G. Dorn,et al.  Cytoplasmic signaling pathways that regulate cardiac hypertrophy. , 2001, Annual review of physiology.

[65]  J. Sadoshima,et al.  The cellular and molecular response of cardiac myocytes to mechanical stress. , 1997, Annual review of physiology.

[66]  M. Pfeffer,et al.  Angiotensin-converting enzyme inhibition and ventricular remodeling after myocardial infarction. , 1995, Annual review of physiology.

[67]  R. Alexander,et al.  Angiotensin II receptor pharmacology. , 1994, Advances in pharmacology.

[68]  H. Brown,et al.  Cardiac pacemaking in the sinoatrial node. , 1993, Physiological reviews.

[69]  G. Booz,et al.  Cardiac actions of angiotensin II: Role of an intracardiac renin-angiotensin system. , 1992, Annual review of physiology.