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Diethylnitrosamine administration in vivo increases hepatic poly(ADP-ribose) levels in rats: results of a modified technique for poly(ADP-ribose) measurement |
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| Authors: | Rawling, Jean M. Driscoll, Eric R. Poirier, Guy G. Kirkland, James B. |
| Affiliation: | 1Department of Nutritional Sciences, University of Guelph Guelph, Ontario, Canada N1G 2W1 1Laboratoire du Metabolisme du poly(ADP-ribose), Endocrinology Moleculaire, Centre Hospitalier de L'Universite Laval 2705 boulevard Laurier, Sainte Foy, Quebec, Canada G1V 4G2 |
| Abstract: | Poly(ADP-ribose) (polymer) is enzymatically synthesized on nuclearproteins in response to DNA strand breaks. NAD+ is the substratefor this reaction, which is catalyzed by Poly(ADP-ribose)polymerase.This post-translational modification occurs in response to DNAstrand breaks and is thought to play an important role in DNArepair. Polymer synthesis resulting from DNA damage has beendescribed in cultured cells, but measurement is more difficultin animal tissues. In this study, modifications were made toan earlier method to measure carcinogen-induced increases inpolymer levels in vivo. RNase I was added to the enzyme mixtureused to digest polymer to ribosyladenosine (RAdo). This preventedthe inhibition of snake venom phosphodiesterase by RNA. TheHPLC analysis was improved, allowing elimination of the secondboronate affinity chromatography step traditionally used topurify |
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