To investigate the functional relevance of a regulatory mutation affecting the enhancer element PEA2 of the rat ANP gene we transfected rat cardiomyocytes and aortic endothelial cells with either the mutant or the wild-type ANP promoter construct (-683 +54) and performed CAT assays both at baseline and in response to Phenylephrine and Angiotensin II. In the myocardial cells we also determined the DNA/nuclear protein interaction through electrophoretic mobility shift assay. These studies showed a significantly lower degree of ANP transcription in the presence of the mutant PEA2 site, thus demonstrating its functional significance and the biological relevance of ANP gene structural alterations.
Effect of a regulatory mutation on the rat atrial natriuretic peptide gene transcription / Rubattu, Speranza Donatella; Giliberti, R; DE PAOLIS, P; Stanzione, R; Spinsanti, Paola; Venturelli, V; Volpe, Massimo. - In: PEPTIDES. - ISSN 0196-9781. - 23(3).:(2002), pp. 555-560. [10.1016/S0196-9781(01)00626-X,]
Effect of a regulatory mutation on the rat atrial natriuretic peptide gene transcription.
RUBATTU, Speranza Donatella;SPINSANTI, Paola;VOLPE, Massimo
2002
Abstract
To investigate the functional relevance of a regulatory mutation affecting the enhancer element PEA2 of the rat ANP gene we transfected rat cardiomyocytes and aortic endothelial cells with either the mutant or the wild-type ANP promoter construct (-683 +54) and performed CAT assays both at baseline and in response to Phenylephrine and Angiotensin II. In the myocardial cells we also determined the DNA/nuclear protein interaction through electrophoretic mobility shift assay. These studies showed a significantly lower degree of ANP transcription in the presence of the mutant PEA2 site, thus demonstrating its functional significance and the biological relevance of ANP gene structural alterations.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
