P53-dependent and -independent links between DNA-damage, apoptosis and mutation frequency in ES cells.

The hypothesis that p53 deficiency enhances the survival of DNA-damage bearing cells was investigated in wild-type and p53 mutant embryonic stem (ES) cells. Following UV-C irradiation, p53 is rapidly induced in wild-type cells and p53-dependent apoptosis follows within 8 h, resulting in the death of the majority of cells within 36 h. Increasing doses of UV-irradiation resulted in enhanced clonogenic survival of null cells as compared to wild-type. Amongst surviving clones, the Hprt mutation frequency was found to be dependent upon UV dose and influenced by p53 status. Treatment with ionizing radiation led to enhanced expression of p53 but resulted in little induction of apoptosis irrespective of p53 status. However, clonogenic potential was considerably reduced, particularly in wild-type cells which showed a tenfold lower survival than null cells. In contrast to the effects of UV-irradiation, the incidence of Hprt mutation did not differ significantly between wild-type and p53 null survivors. The data confirm that p53 restricts the numbers of cells bearing mutations that survive DNA damage induced by either agent, albeit by different mechanisms.