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Departments of Medicine and Biochemistry and Cancer Center, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4937
We have used two different approaches to study the consequences of NAD/poly(ADP-ribose) deficiency on p53 expression and its activity in V79-derived cell lines. In the first approach, we have used two cell lines that are deficient in poly(ADP-ribose) (pADPR) synthesis because of deficiency in the enzyme poly(ADP-ribose) polymerase (PARP). In a second approach, we have used a cell line that is deficient in NAD/pADPR metabolism due to unavailability of NAD, the substrate for PARP. These NAD/PARP-deficient cell lines exhibit a significant reduction in both baseline p53 expression and its activity compared to their parental V79 cells. Furthermore, etoposide, a topoisomerase II inhibitor that was shown to cause an increase in p53 expression and subsequent apoptosis in V79 cells, failed to produce any significant increase in p53 expression or apoptotic DNA fragmentation in NAD/PARP-deficient cell lines. Thus, our studies suggest that NAD/pADPR synthesis may be involved in the regulation of p53 and its dependent pathways.
1 This work was supported in part by American Cancer Society Grant IRG-186 and National Cancer Institute Grants PO1-CA48735, PO1-CA51183, and P3O-CA43703.
2 To whom requests for reprints should be addressed, at Case Western Reserve University, School of Medicine, BRB 301A, 10900 Euclid Avenue, Cleveland OH 44106-4937.
Received 5/30/95. Accepted 7/20/95.
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