Growth inhibition and cell killing by N-methyl-N-nitrosourea: metabolic alterations that accompany poly(ADP-ribosyl)ation.
The effect of inhibition of poly(ADP-ribose) polymerase (PARP) on the growth arrest and cell killing induced by N-methyl-N-nitrosourea (MNU) was studied in L929 fibroblasts. Depletion of NAD and ATP preceded the cell killing by a 1-h exposure to 10 or 15 mM MNU. 3-Aminobenzamide (ABA), an inhibitor of PARP, spared the depletion of NAD and ATP and prevented the cell killing. With 5 mM MNU, a depletion of NAD was promptly reversed, and there was no loss of ATP and no cell death. Aphidicolin, a DNA polymerase inhibitor, prevented the restoration of NAD, with resulting depletion of ATP and death of the cells, effects that were prevented by ABA. Azide together with 2-deoxyglucose depleted ATP, followed by a loss of NAD and cell death, changes that occurred in the absence of DNA single strand breaks (DNA SSB). ABA prevented the depletion of NAD, but not that of ATP, nor the cell killing. MNU (2.5 mM) inhibited cell growth without effect on the viability of the cells. ABA potentiated the cell growth inhibition. Thus, inhibition of PARP potentiates cell growth inhibition by limiting DNA repair mechanisms. Alternatively, inhibition of the DNA repair response to more extensive DNA damage prevents cell killing. The ATP depletion caused by poly(ADP-ribosyl)ation, rather than DNA SSB and the loss of NAD, is the more critical event in the cell killing.
Mizumoto
K
K
Department of Pathology, Thomas Jefferson University, Phildelphia, Pennsylvania 19107, USA.
Farber
J L
JL
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D001386:Azides / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003847:Deoxyglucose / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects; D008770:Methylnitrosourea / Q000633:toxicity*; D051379:Mice; D009243:NAD / Q000032:analysis; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism* / Q000494:pharmacology; D019810:Sodium Azide
D001386:Azides; D009243:NAD; D016590:Aphidicolin; D008770:Methylnitrosourea; D000255:Adenosine Triphosphate; D019810:Sodium Azide; D003847:Deoxyglucose; D011065:Poly(ADP-ribose) Polymerases