Elevated levels of wild-type p53 induced by radiolabeling of cells leads to apoptosis or sustained growth arrest.
BACKGROUND
The tumor suppressor protein p53 regulates progression through the checkpoint between the G1 and S phases of the cell cycle in response to radiation- or drug-induced DNA damage. We have examined potential p53-mediated effects of metabolically labeling cultured mammalian cells with [35S]methionine and [3H]thymidine, methods that are commonly used to study the biochemical properties, synthesis, processing and degradation of proteins and the replication of DNA in proliferating cells.


RESULTS
Wild-type p53 protein concentrations rapidly increased to high levels following metabolic radiolabeling of cells, as determined by four distinct assays. The increased concentration of wild-type p53 resulted in apoptosis of normal human peripheral blood lymphocytes and of murine T-cell acute lymphoblastic leukemia cells. In leukemia cells containing no p53, or only mutant p53 alleles, p53 protein levels were not increased and the cells did not undergo apoptosis in response to radioactive labeling. Radiolabeling of human diploid fibroblasts resulted in a prolonged growth arrest that was maintained for nearly three weeks.


CONCLUSIONS
The results of experiments employing radiolabeling techniques to characterize various aspects of cellular physiology may be seriously influenced by the induction of aberrant cell-cycle arrest and/or apoptosis mediated by wild-type p53. Furthermore, our observations suggest that stabilization of wild-type p53 in response to irradiation may not act primarily to facilitate the repair of DNA damage by inducing a transient G1-phase arrest, but rather to ensure genetic stability through sustained cell-cycle arrest or apoptotic death of the damaged cells.
Yeargin
J
J
Department of Biology, University of California, San Diego, La Jolla 92093-0063, USA.
Haas
M
M
D000818:Animals; D017209:Apoptosis / Q000528:radiation effects*; D002455:Cell Division / Q000528:radiation effects*; D002460:Cell Line; D002478:Cells, Cultured; D006801:Humans; D007553:Isotope Labeling; D007963:Leukocytes, Mononuclear / Q000378:metabolism / Q000528:radiation effects; D008715:Methionine / Q000378:metabolism; D051379:Mice; D010761:Phosphorus Radioisotopes / Q000494:pharmacology*; D011233:Precipitin Tests; D013936:Thymidine / Q000378:metabolism; D014316:Tritium / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism / Q000528:radiation effects*
D010761:Phosphorus Radioisotopes; D016159:Tumor Suppressor Protein p53; D014316:Tritium; D008715:Methionine; D013936:Thymidine