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The expression of endothelin-1 (ET-1) in cardiac myocytes is markedly induced during the development of heart failure in vivo and by stimulation with the alpha(1)-adrenergic agonist phenylephrine in culture. Although recent studies have suggested a role for cardiac-specific zinc finger GATA factors in the transcriptional pathways that modulate cardiac hypertrophy, it is unknown whether these factors are also involved in cardiac ET-1 transcription and if so, how these factors are modulated during this process. Using transient transfection assays in primary cardiac myocytes from neonatal rats, we show here that the GATA element in the rat ET-1 promoter was required for phenylephrine-stimulated ET-1 transcription. Cardiac GATA-4 bound the ET-1 GATA element and activated the ET-1 promoter in a sequence-specific manner. Stimulation by phenylephrine caused serine phosphorylation of GATA-4 and increased its ability to bind the ET-1 GATA element. Inhibition of the extracellularly responsive kinase cascade with PD098059 blocked the phenylephrine-induced increase in the DNA binding ability and the phosphorylation of GATA-4. These findings demonstrate that serine phosphorylation of GATA-4 is involved in alpha(1)-adrenergic agonist-responsive transcription of the ET-1 gene in cardiac myocytes and that extracellularly responsive kinase 1/2 activation plays a role upstream of GATA-4.

Leukemia inhibitory factor is a member of a family of structurally related cytokines sharing the receptor component gp130. Activation of gp130 by leukemia inhibitory factor is sufficient to induce myocardial cell hypertrophy accompanied by specific changes in the pattern of gene expression. However, the molecular mechanisms that link gp130 activation to these changes have not been clarified. The present study investigated the transcriptional pathways by which leukemia inhibitory factor activates beta-myosin heavy chain expression during myocardial cell hypertrophy. Mutation of the GATA motif in the beta-myosin heavy chain promoter totally abolished leukemia inhibitory factor-responsive transcription without changing basal transcriptional activity. In contrast, endothelin-1 responsiveness was unaffected by the GATA mutation. Among members of the cardiac GATA transcription factor subfamily (GATA-4, -5, and -6), GATA-5 was the sole and potent transactivator for the beta-myosin heavy chain promoter. This transactivation was dependent on sequence-specific binding of GATA-5 to the beta-myosin heavy chain GATA element. Cardiac nuclear factors that bind to to the beta-myosin heavy chain GATA element were induced by leukemia inhibitory factor stimulation. Last, leukemia inhibitory factor stimulation markedly increased transcripts of cardiac GATA-5, the expression of which is normally restricted to the early embryo. Thus, GATA-5 may be involved in gp130 signaling in cardiac myocytes.

A cellular target of adenovirus E1A oncoprotein, p300 is a transcriptional coactivator and a negative regulator of cellular proliferation. A previous study suggests that the p300 family is also involved in cell type-specific transcription in cardiac myocytes. However, nothing is known about which cardiac transcription factor(s) interact with and transactivate through these proteins. The transcription factors GATA-4/5/6 have been implicated as key regulators of cardiogenesis, and they participate in the transcription of many cardiac-specific genes. Here we show that E1A represses the GATA-5-dependent transactivation of a promoter derived from the cardiac-restricted atrial natriuretic factor gene. This repression is correlated with the interaction of E1A with p300, indicating that p300 participates in GATA-5-dependent transactivation. E1A markedly down-regulates endogenous atrial natriuretic factor expression, as well as disrupts the interaction between p300 and GATA-5. A small fragment of p300 containing the carboxyl-terminal cysteine/histidine-rich domain, sufficient to interact with GATA-5, prevents transcriptional activation by GATA-5 as a dominant-negative mutant. Consistent with its role as a coactivator, p300 markedly potentiates GATA-5-activated transcription. These results implicate p300 as an important component of myocardial cell differentiation and provide an insight into the relationship between mechanisms that mediate cell type-specific transcription and cell cycle regulation during cardiogenesis.

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Two topics related to the FEM simulation of multi-step stamping processes are discussed. They are: (1) the difficulty in simulating the springback and trimming operations, and (2) the way to select a most appropriate element. The basic nature of problems with the springback simulation can be attributed to a ‘drift’ from the equilibrium path due to the static explicit formulation of the ITAS3D FEM code used for the simulation. A strategy on how to minimize the ‘drift’ phenomenon is presented here. The second problem of the most suitable element, shell or solid, for simulation of multi-step stamping processes is of general character and can be addressed to all users of explicit and implicit FEM codes. We show that for some of the stamping processes, a solid element is superior to a shell. Finally, the entire simulation of multi-step forming processes of a car wheel disk is presented and compared with experiment data.

Two topics related to the FEM simulation of multi-step stamping processes are discussed. They are: (1) the difficulty in simulating the springback and trimming operations, and (2) the way to select a most appropriate element. The basic nature of problems with the springback simulation can be attributed to a ‘drift’ from the equilibrium path due to the static explicit formulation of the ITAS3D FEM code used for the simulation. A strategy on how to minimize the ‘drift’ phenomenon is presented here. The second problem of the most suitable element, shell or solid, for simulation of multi-step stamping processes is of general character and can be addressed to all users of explicit and implicit FEM codes. We show that for some of the stamping processes, a solid element is superior to a shell. Finally, the entire simulation of multi-step forming processes of a car wheel disk is presented and compared with experiment data.

The adenylate cyclase of a human medullary thyroid carcinoma was activated by TRH, glucagon, epinephrine, norepinephrine, phentolamine, serotonin and NaF, suggesting the presence of multiple hormone receptors including a β-adrenergic one in the tumor.