DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways.

Abstract

The tumor suppressor protein p53 serves as a critical regulator of a G1 cell cycle checkpoint and of apoptosis following exposure of cells to DNA-damaging agents. The mechanism by which DNA-damaging agents elevate p53 protein levels to trigger G1/S arrest or cell death remains to be elucidated. In fact, whether damage to the DNA template itself participates in transducing the signal leading to p53 induction has not yet been demonstrated. We exposed human cell lines containing wild-type p53 alleles to several different DNA-damaging agents and found that agents which rapidly induce DNA strand breaks, such as ionizing radiation, bleomycin, and DNA topoisomerase-targeted drugs, rapidly triggered p53 protein elevations. In addition, we determined that camptothecin-stimulated trapping of topoisomerase I-DNA complexes was not sufficient to elevate p53 protein levels; rather, replication-associated DNA strand breaks were required. Furthermore, treatment of cells with the antimetabolite N(phosphonoacetyl)-L-aspartate (PALA) did not cause rapid p53 protein increases but resulted in delayed increases in p53 protein levels temporally correlated with the appearance of DNA strand breaks. Finally, we concluded that DNA strand breaks were sufficient for initiating p53-dependent signal transduction after finding that introduction of nucleases into cells by electroporation stimulated rapid p53 protein elevations. While DNA strand breaks appeared to be capable of triggering p53 induction, DNA lesions other than strand breaks did not. Exposure of normal cells and excision repair-deficient xeroderma pigmentosum cells to low doses of UV light, under conditions in which thymine dimers appear but DNA replication-associated strand breaks were prevented, resulted in p53 induction attributable to DNA strand breaks associated with excision repair. Our data indicate that DNA strand breaks are sufficient and probably necessary for p53 induction in cells with wild-type p53 alleles exposed to DNA-damaging agents.

Comparative alterations in p53 expression and apoptosis in the irradiated rat small and large intestine

“…Alterations in p53 protein levels were very rapid, peaking within 4 h in both the small and large intestine, and then reducing to approximately normal levels at 24 h. In comparison with our results using X-ray irradiation, increased p53 protein levels were reported to be detectable for at least 20 days after UV radiation (Fritsche et al, 1993;Hall et al, 1993;Lee and Bernstein, 1993;Lu and Lane, 1993;Nelson and Kastan, 1994). It is possible that the role of p53 protein in these two examples differs, or that the stimulus for p53 expression is important in conferring different types of p53 protein stability.…”

Identification of a novel p53-dependent activation pathway of STAT1 by antitumour genotoxic agents

“…We first used actinomycin D, a well-known agent able to both block mRNA synthesis and to induce genotoxic stress and p53 activation. 18 We found that actinomycin D induced Y701-STAT1 phosphorylation after 1 h treatment (Figure 2a). Meanwhile, mRNA levels of p21, a target gene of both p53 and STAT1, were significantly decreased after actinomycin D treatment (Figure 2b), in agreement with inhibition of mRNA synthesis by this agent.…”

Tracking the cell cycle origins for escape from topotecan action by breast cancer cells

“…The activation of the tumor suppressor protein p53 plays an important, though not essential, role in the activation of the responses of the genotoxic stress pathways (Schartz and Rotter, 1998). The activation of p53 may even provide a means to distinguish the consequences of genotoxic dsDNA break induction from less-toxic ssDNA break formation (Nelson and Kastan, 1994). The elevation of p53 protein levels was detected, by flow cytometry, using the DO-1 antibody, immediately following a 1 h exposure to topotecan.…”