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Stimulation of Polyadenosine Diphosphoribose Synthesis by DNA Lesions Induced by Sodium Chromate in Chinese Hamster V-79 Cells¹

Department of Medical Biochemistry [M. S., T. N., T. H., R. O.], Kurume University School of Medicine, 67 Asahi-machi, Kurume 830, Japan, and Laboratory of Molecular Toxicology and Chemical Carcinogenesis, Institute of Environmental Medicine [M. C.], New York University Medical Center, 550 First Avenue, New York, New York 10016

Polyadenosine diphosphoribose [poly(ADP-ribose)] synthesis was stimulated by DNA lesions induced with Na₂CrO₄ and methyl methanesulfonate (MMS) in Chinese hamster V-79 cells. Na₂CrO₄ and MMS induced DNA single-strand breaks in a concentration-dependent manner; however, the breaks induced by Na₂CrO₄ were "protein associated" while those induced by MMS were not. MMS stimulated in a dose-dependent fashion the synthesis of poly(ADP-ribose) up to 6-fold above the control. Na₂CrO₄ also induced poly(ADP-ribose) synthesis, but the level of synthesis was less than 3-fold. Control experiments demonstrated that Na₂CrO₄ treatment of cells did not affect their ability to synthesize poly(ADP-ribose) in response to DNA damage. Treatment of cells with Na₂CrO₄ and MMS induced more poly(ADP-ribose) synthesis than each agent alone; however, whenever Na₂CrO₄ was utilized, the breaks required proteinase K to be detected. Following removal of extracellular chromate, the DNA strand breaks induced by 0.2 mM Na₂CrO₄ were repaired quickly during the first hour but more slowly for the next 3 h. In the presence of 3-aminobenzamide, an inhibitor of poly(ADP-ribose) synthesis, the repair of DNA breaks was reduced. These results suggest that DNA protein-associated breaks produced by Na₂CrO₄ were recognized by poly(ADP-ribose) polymerase and that there are differences in poly(ADP-ribose) synthesis in response to Na₂CrO₄ and MMS. The results also suggest that the repair of breaks induced by Na₂CrO₄ are associated with poly(ADP-ribose) synthesis, but perhaps because most of these breaks are protein associated, there is less stimulation of poly(ADP-ribose) synthesis.

¹ Supported by Grant CA43070 of the National Cancer Institute and Grant R813140-010 of the United States Environmental Protection Agency.

² To whom requests for reprints should be addressed, at Department of Medical Biochemistry, Kurume University School of Medicine, 67 Asahi-machi, Kurume 830, Japan.

Received 6/17/87. Revised 9/ 9/87. Revised 11/18/87. Accepted 11/20/87.