Oxidative stress enhances phosphorylation of p53 in neonatal rat cardiomyocytes.
p53 is an important regulator of cell growth and apoptosis and its activity is regulated by phosphorylation. Accordingly, in neonatal rat cardiomyocytes we examined the involvement of p53 in H(2)O(2)-induced apoptosis. Treatment with 50-100 microM H(2)O(2) markedly induced apoptosis in cardiomyocytes, as assessed by gel electrophoresis of genomic DNA. To examine whether H(2)O(2) increases p53 phosphorylation in cardiomyocytes, we utilized an antibody that specifically recognizes phosphorylated p53 at serine-15. The level of phosphorylated p53 was markedly increased by 100 microM H(2)O(2) at 30 and 60 min. Using specific protein kinase inhibitors we examined the involvement of protein kinases in p53 phosphorylation in response to H(2)O(2) treatment. However, staurosporine, a broad spectrum inhibitor of protein kinases, SB202190, a specific p38 kinase inhibitor, PD98059, a MAP kinase inhibitor, wortmannin, an inhibitor of DNA-PK and PI3 kinase, SP600125, a JNK inhibitor and caffeine,an inhibitor of ATM and ATR, failed to prevent the H(2)O(2)-induced phosphorylation of p53. cDNA microarray revealed that H(2)O(2) markedly increased expression of several p53 upstream modifiers such as the p300 coactivator protein and several downstream effectors such as gadd45, but decreased the expression of MDM2, a negative regulator of p53. Our results suggest that phosphorylation of p53 at serine-15 may be an important signaling event in the H(2)O(2)-mediated apoptotic process.
Long
Xilin
X
The Molecular Cardiology and Neuromuscular Institute, Highland Park, NJ 08904, USA.
Goldenthal
Michael J
MJ
Marín-García
José
J
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D006861:Hydrogen Peroxide / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D010766:Phosphorylation; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
D016877:Oxidants; D016159:Tumor Suppressor Protein p53; D006861:Hydrogen Peroxide; D020928:Mitogen-Activated Protein Kinases