Effect of a regulatory mutation on the rat atrial natriuretic peptide gene transcription.
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.
Rubattu
Speranza
S
Istituto Neuromed, Polo Molisano dell'Universita' La Sapienza di Roma, Pozzilli, Italy. rubattu.speranza@neuromed.it
Giliberti
Rosangela
R
De Paolis
Paola
P
Stanzione
Rosita
R
Spinsanti
Paola
P
Venturelli
Vanessa
V
Volpe
Massimo
M
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D009320:Atrial Natriuretic Factor