Myocyte-Specific M-CAT and MEF-1 Elements Regulate G-Protein Gamma 3 Gene (γ 3 ) Expression in Cardiac Myocytes

Abstract

Little is known regarding the mechanisms that control the expression of G-protein α, β, and γ subtypes. We have previously shown that the G-protein γ ₃ gene is expressed in the heart, brain, lung, spleen, kidney, muscle, and testis in mice. We have also reported that the G-protein γ ₃ subunit is expressed in rat cardiac myocytes, but not in cardiac fibroblasts. Other studies have shown that the γ ₃ subunit couples to the angiotensin A1A receptor in portal vein myocytes, and has been shown to mediate β-adrenergic desensitization in cardiac myocytes treated with atorvastatin. In the present study, we evaluated G-protein γ ₃ promoter-luciferase reporter constructs in primary myocytes to identify key regulatory promoter regions. We identified two important regions of the promoter (upstream promoter region [UPR] and downstream promoter region [DPR]), which are required for expression in cardiac myocytes. We observed that removal of 48 bp in the UPR diminished gene transcription by 75%, and that the UPR contains consensus elements for myocyte-specific M-CAT and myocyte enhancer factor 1 (MEF-1) elements. The UPR and DPR share transcription factor elements for myocyte-specific M-CAT element. We observed that cardiac myocyte proteins bind to γ ₃ oligonucleotides containing transcription factor elements for myocyte-specific M-CAT and MEF-1. Myocyte-specific M-CAT proteins were supershifted with transcriptional enhancer factor-1 (TEF-1) antibodies binding to the γ ₃ M-CAT element, which is in agreement with reports showing that the M-CAT element binds the TEF-1 family of transcription factors. The 150 bp DPR contains three M-CAT elements, an INR element, an upstream stimulatory factor 1 element, and the transcription start site. We have shown that myocyte γ ₃ gene expression is regulated by myocyte-specific M-CAT and MEF-1 elements.