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DNA Methylation Inhibitor Regulates Guanylyl Cyclase/Natriuretic Peptide Receptor‐A Gene Expression in Mesangial Cells
Author(s) -
Kumar Prerna,
Pandey Kailash
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.880.25
Subject(s) - npr1 , microbiology and biotechnology , dna methylation , npr2 , gene expression , chemistry , biology , natriuretic peptide , gene , medicine , biochemistry , heart failure
Cardiac hormone atrial natriuretic peptide binds to guanylyl cyclase/natriuretic peptide receptor‐A (GC‐A/NPRA) and produces second messenger cGMP, which plays a pivotal role in maintaining blood pressure and cardiovascular homeostasis. The present study was aimed at understanding the mechanisms involved in DNA methylation inhibitor 5‐Azacytidine (5‐aza)‐mediated Npr1 (coding for GC‐A/NPRA) gene transcription. The studies were carried out in mouse mesangial cells (MMCs) cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum and ITS (insulin, transferrin, and sodium selenite) and treated with 5‐aza for 24 h. The MethPrimer search result showed the presence of three cytosine‐phosphate‐guanine (CpG) islands on the Npr1 full length promoter, namely island 1 (‐886 to ‐752, 134 bp), island 2 (‐310 to ‐58, 152 bp), and island 3 (‐514 to +310, 464 bp) from the transcription start site. Bisulfite‐Sequencing PCR showed the proximal promoter region ‐356 to +55 of the Npr1 promoter is mostly unmethylated in MMCs. 5‐azacytidine induced mRNA levels of Npr1 gene by 5‐fold and NPRA protein expression by 3‐fold in MMCs in a dose‐dependent manner. Treatment of MMCs with 5‐aza significantly increased acetylation levels of histones H3K9/14 and H4K12 (p<0.001). The results demonstrate that 5‐aza regulates Npr1 gene expression by modulation of DNA methylation and histone modifications. The present findings provide a novel regulatory mechanism for Npr1 gene transcription, an important player in the control of hypertension and cardiovascular homeostasis.