PUTATIVE ROLE FOR NICOTINAMIDE ADENINE DINUCLEOTIDE-PROMOTED NUCLEAR PROTEIN MODIFICATION IN ANTITUMOR ACTIVITY OF N-METHYL-N-NITROSOUREA

Abstract

Incubation of HeLa cells with the anticancer agent N-methyl-N-nitrosourea (MNU) results in: depression of intracellular NAD levels; stimulation of the chromatin-associated, chromosomal protein-modifying enzyme polyadenosine diphosphoribose [poly(ADP-ribose)] polymerase, which uses NAD as substrate; and some fragmentation of cellular DNA. DNase treatment of HeLa nuclei in vitro also stimulates poly(ADP-ribose) polymerase activity, but not in nuclei derived from MNU-treated cells unless they have been subsequently incubated to allow for recovery from MNU damage. DNA polymerase activity is stimulated in vitro by poly(ADP) ribosylation of nuclear proteins. By using intact nuclei derived from MNU-treated HeLa cells, the repair via elongation of single-strand DNA breaks is demonstrated im vitro. This repair is dependent on DNA polymerase activity and is enhanced by ADP ribosylation of histones. Inhibition of poly(ADP-ribose) polymerase with nicotinamide results in extensive degradation of MNU-damaged DNA. Taken as a whole, these results suggest that poly(ADP-ribose) polymerase may play a role in the repair of alkylation damage to cellular DNA and that the inhibition of this enzyme in vivo might be exploited to potentiate the antitumor and carcinogenic activities of MNU.