title: Poly(ADP-ribose) polymerase-1-dependent cardiac myocyte cell death during heart failure is mediated by NAD+ depletion and reduced Sir2alpha deacetylase activity.
abstract: Robust activation of poly(ADP-ribose) polymerase-1 (PARP) by oxidative stress has been implicated as a major cause of caspase-independent myocyte cell death contributing to heart failure. Here, we show that depletion of myocyte NAD levels and the subsequent reduction of Sir2alpha deacetylase activity are the sequential steps contributing to PARP-mediated myocyte cell death. In both failing hearts and cultured cardiac myocytes, the increased activity of PARP was associated with depletion of cellular NAD levels and reduced Sir2alpha deacetylase activity. Myocyte cell death induced by PARP activation was prevented by repletion of cellular NAD levels either by adding NAD directly to the culture medium or by overexpressing NAD biosynthetic enzymes. The beneficial effect of NAD repletion was seen, however, only when Sir2alpha was intact. Knocking down Sir2alpha levels by small interfering RNA eliminated this benefit, indicating that Sir2alpha is a downstream target of NAD replenishment leading to cell protection. NAD repletion also prevented loss of the transcriptional regulatory activity of the Sir2alpha catalytic core domain resulting from PARP activation. We also show that PARP activation and the concomitant reduction of Sir2alpha activity in failing hearts regulate the post-translational acetylation of p53. These data demonstrate that, in stressed cardiac myocytes, depletion of cellular NAD levels forms a link between PARP activation and reduced Sir2alpha deacetylase activity, contributing to myocyte cell death during heart failure.
authors.affiliation: Department of Cardiothoracic Surgery, University of Chicago, IL 60637, USA.
authors.forename: Jyothish B
authors.initials: JB
authors.lastname: Pillai
authors.forename: Ayman
authors.initials: A
authors.lastname: Isbatan
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Imai
authors.forename: Mahesh P
authors.initials: MP
authors.lastname: Gupta
mesh_terms: D000818:Animals; D001011:Aorta / Q000473:pathology; D015153:Blotting, Western; D019556:COS Cells; D002118:Calcium / Q000378:metabolism; D020134:Catalytic Domain; D016923:Cell Death; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004247:DNA / Q000378:metabolism; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D006333:Heart Failure / Q000473:pathology; D006352:Heart Ventricles / Q000473:pathology; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D008954:Models, Biological; D032342:Muscle Cells / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000473:pathology*; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D018384:Oxidative Stress; D010957:Plasmids / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D056564:Sirtuin 1; D037761:Sirtuins / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D009243:NAD; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases; C447939:SIRT1 protein, human; C488368:Sirt1 protein, rat; D056564:Sirtuin 1; D037761:Sirtuins; D006655:Histone Deacetylases; D002118:Calcium