Inactivation of the poly(ADP-ribose) polymerase gene affects oxygen radical and nitric oxide toxicity in islet cells.
Activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) is an early response of cells exposed to DNA-damaging compounds such as nitric oxide (NO) or reactive oxygen intermediates (ROI). Excessive poly-(ADP-ribose) formation by PARP has been assumed to deplete cellular NAD+ pools and to induce the death of several cell types, including the loss of insulin-producing islet cells in type I diabetes. In the present study we used cells from mice with a disrupted and thus inactivated PARP gene to provide direct evidence for a causal relationship between PARP activation, NAD+ depletion, and cell death. We found that mutant islet cells do not show NAD+ depletion after exposure to DNA-damaging radicals and are more resistant to the toxicity of both NO and ROI. These findings directly prove that PARP activation is responsible for most of the loss of NAD+ following such treatment. The ADP-ribosylation inhibitor 3-aminobenzamide partially protected islet cells with intact PARP gene but not mutant cells from lysis following either NO or ROI treatment. Hence the protective action of 3-aminobenzamide must be due to inhibition of PARP and does not result from its other pharmacological properties such as oxygen radical scavenging. Finally, the use of mutant cells an alternative pathway of cell death was discovered which does not require PARP activation and NAD+ depletion. In conclusion, the data prove the causal relationship of PARP activation and subsequent islet cell death and demonstrate the existence of an alternative pathway of cell death independent of PARP activation and NAD+ depletion.
Heller
B
B
Diabetes Research Institute, University of Düsseldorf, Federal Republic of Germany.
Wang
Z Q
ZQ
Wagner
E F
EF
Radons
J
J
Bürkle
A
A
Fehsel
K
K
Burkart
V
V
Kolb
H
H
D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D002467:Cell Nucleus / Q000201:enzymology; D002478:Cells, Cultured; D004249:DNA Damage; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D007700:Kinetics; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D009243:NAD / Q000378:metabolism; D009569:Nitric Oxide / Q000633:toxicity*; D009599:Nitroprusside / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000096:biosynthesis / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000633:toxicity*; D014969:Xanthine Oxidase / Q000494:pharmacology
D001549:Benzamides; D017382:Reactive Oxygen Species; D009243:NAD; D009599:Nitroprusside; D009569:Nitric Oxide; C025160:3-aminobenzamide; D014969:Xanthine Oxidase; D011065:Poly(ADP-ribose) Polymerases