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| Item Type: | Article |
|---|---|
| Title: | Identification of a cis-acting regulatory element conferring inducibility of the atrial natriuretic factor gene in acute pressure overload |
| Creators Name: | von Harsdorf, R., Edwards, J.G., Shen, Y.T., Kudej, R.K., Dietz, R., Leinwand, L.A., Nadal-Ginard, B. and Vatner, S.F. |
| Abstract: | To identify the cis-acting regulatory element(s) which control the induction of the atrial natriuretic factor (ANF) gene in acute pressure overload, DNA constructs consisting of promoter elements linked to a reporter gene were injected into the myocardium of dogs, which underwent aortic banding or were sham-operated. Expression of a reporter gene construct harboring the ANF promoter (-3400ANF) was induced 6-12-fold after 7 d of pressure overload. An internal deletion of 556 bp (nucleotide sequence -693 to -137) completely abrogated the inducibility of the ANF reporter gene construct. An activator protein-1 (AP1)-like site (-496 to -489) and a cAMP regulatory element (CRE) (-602 to -596) are located within the deleted sequence. Site-directed mutagenesis of the AP1-like site but not the CRE completely prevented the induction of this construct to acute pressure overload. Further, the AP1-like site was able to confer inducibility of a heterologous promoter (beta-myosin heavy chain) to higher values than controls. Gel mobility shift assay (GMSA) supershift analysis was performed using a radiolabeled probe of the ANF promoter (-506/-483) that included the AP1-like site (ATGAATCA) sequence, as well as a probe converted to contain an AP1 consensus sequence (ATGACTCA). GMSA analysis demonstrated that the ANF AP1-like element could bind both a constitutively expressed factor and the AP1 proteins, and conversion to a true AP1 site increased its affinity for AP1. However, 7 d after the onset of pressure overload, the AP1 proteins were present only at low levels, and the major complex formed by the ANF AP1-like probe was not supershifted by a jun antibody. Using a large animal model of pressure overload, we have demonstrated that a unique cis-acting element was primarily responsible for the overload induction of the ANF gene. |
| Keywords: | Atrial Natriuretic Factor, Cardiomegaly, DNA-Binding Proteins, Gene Expression Regulation, Regulator Genes, Promoter Regions (Genetics), Atrial Natriuretic Factor Receptors, Trans-Activation (Genetics), Transcription Factor AP-1, Animals, Dogs |
| Source: | Journal of Clinical Investigation |
| ISSN: | 0021-9738 |
| Publisher: | American Society for Clinical Investigation |
| Volume: | 100 |
| Number: | 5 |
| Page Range: | 1294-1304 |
| Date: | 1 September 1997 |
| Official Publication: | https://doi.org/10.1172/JCI119643 |
| PubMed: | View item in PubMed |
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