Poly(ADP-ribose) turnover in quail myoblast cells: relation between the polymer level and its catabolism by glycohydrolase.
The concerted action of poly(ADP-ribose) polymerase (PARP) which synthesizes the poly(ADP-ribose) (pADPr) in response to DNA strand breaks and the catabolic enzyme poly(ADP-ribose) glycohydrolase (PARG) determine the level of polymer and the rate of its turnover. In the present study, we have shown that the quail myoblast cells have high levels of basal polymer as compared to the murine C3H10T1/2 fibroblasts. We have conducted this study to investigate how such differences influence polymer synthesis and its catabolism in the cells in response to DNA damage by alkylating agent. In quail myoblast cells, the presence of high MNNG concentration such as 200 microM for 30 min induced a marginal decrease of 15% in the NAD content. For C3H10T1/2 cell line, 64 microM MNNG provoked a depletion of NAD content by approximately 50%. The induction of the polymer synthesis in response to MNNG treatment was 6-fold higher in C3H10T1/2 cells than in quail myoblast cells notwithstanding the fact that 3-fold higher MNNG concentration was used for quail cells. The polymer synthesis thus induced in quail myoblast cells had a 4-5 fold longer half life than those induced in C3H10T1/2 cells. To account for the slow turnover of the polymer in the quail myoblast cells, we compared the activities of the polymer catabolizing enzyme (PARG) in the two cell types. The quail myoblast cells had about 25% less activity of PARG than the murine cells. This difference in activity is not sufficient to explain the large difference of the rate of catabolism between the two cell types implicating other cellular mechanisms in the regulation of pADPr turnover.
Affar
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Health and Environment Unit, CHUL Research Center, CHUQ, Ste-Foy, Québec, Canada.
Shah
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Poirier
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D000818:Animals; D002460:Cell Line; D003370:Coturnix; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007700:Kinetics; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D051379:Mice; D009132:Muscles / Q000378:metabolism*; D009153:Mutagens / Q000494:pharmacology; D009243:NAD / Q000032:analysis; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis / Q000378:metabolism*; D013997:Time Factors
D009153:Mutagens; D009243:NAD; D008769:Methylnitronitrosoguanidine; D011064:Poly Adenosine Diphosphate Ribose; D006026:Glycoside Hydrolases; C022407:poly ADP-ribose glycohydrolase