α 1-Adrenergic activation of the cardiac ankyrin repeat protein gene in cardiac myocytes

Abstract

Cardiac ankyrin repeat protein (CARP) is a nuclear transcription cofactor that is activated by multiple signaling pathways in hypertrophic cardiac myocytes. Since CARP has been reported to be a transcriptional co-repressor, its activation during hypertrophy might contribute to the deregulation of gene expression leading to heart failure. Here, we found that α 1-adrenergic signaling activates CARP mRNA expression in rat cardiac myocytes. To examine how α 1-adrenergic signaling activates the CARP gene, a 660 bp fragment of the mouse CARP promoter was cloned. Previous reports suggested that the mouse CARP promoter was dependent on the GATA4 transcription factor whereas the human CARP promoter was dependent on transcriptional enhancer factor-1 (TEF-1). TEF-1 and GATA4 transcription factors, known mediators of α 1-adrenergic signaling, bound to the mouse CARP promoter at several sites as determined by gel mobility shift assays. These sites are highly conserved between the mouse and human promoters, suggesting that they are functionally important in both. Mutation analysis showed that binding of TEF-1 factors is required for basal activity of the CARP promoter in cardiac myocytes. However, over-expression of TEF-1 factors could not potentiate the response of the CARP promoter to α 1-adrenergic stimulation. On the other hand, the α 1-adrenergic response was potentiated by GATA4 over-expression. Taken together, our results demonstrate that α 1-adrenergic signaling regulates CARP expression in cardiac myocytes, in part through the transcription factor GATA4.