p53 modulates base excision repair activity in a cell cycle-specific manner after genotoxic stress.
To elucidate the nature of the cross-talk between the p53 protein and the DNA repair machinery, we have investigated the relationship between the two throughout the cell cycle. Base excision repair (BER) was analyzed in cell cycle phase-enriched populations of lymphoid cells expressing wild-type p53. Our study yielded the following novel findings: (a) BER exhibited two distinct peaks of activity, one associated with the G0-G1 checkpoint and the second with the G2-M checkpoint; (b) although the overall BER activity was reduced after exposure of cells to 400R, there was an augmentation of the G0-G1-associated BER activity and a reduction in the G2-M-associated BER activity; and (c) modulations in these patterns of BER after genotoxic stress were found to be p53 regulated. p53 protein levels induced after gamma-irradiation were distributed evenly in the various cell cycle populations (analyzed by the PAb-248 anti-p53 monoclonal antibody). However, both the dephosphorylation of serine 376 of p53 (contained in the PAb-421 epitope) and the specific DNA binding activity, as well as apoptosis, were enhanced toward the G2-M populations. Furthermore, inactivation of wild-type p53, mediated by mutant p53 expression, abolished the alterations in the BER pattern and showed no induction of a G2-M-associated apoptosis after gamma-irradiation. These results suggest that after genotoxic stress, stabilized p53 enhances the G0-G1-associated BER activity, whereas it predominantly reduces BER activity at the G2-M-enriched populations and instead induces apoptosis. After genotoxic stress, p53 functions as a modulator that determines the pattern of BER activity and apoptosis in a cell cycle-specific manner.
Offer
H
H
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Zurer
I
I
Banfalvi
G
G
Reha'k
M
M
Falcovitz
A
A
Milyavsky
M
M
Goldfinger
N
N
Rotter
V
V
D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000528:radiation effects; D001402:B-Lymphocytes / Q000166:cytology / Q000528:radiation effects; D002453:Cell Cycle / Q000502:physiology* / Q000528:radiation effects; D002455:Cell Division / Q000235:genetics / Q000528:radiation effects; D002460:Cell Line / Q000166:cytology / Q000528:radiation effects; D002469:Cell Separation; D002498:Centrifugation; D004247:DNA / Q000096:biosynthesis; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics / Q000502:physiology* / Q000528:radiation effects; D005720:Gamma Rays; D007399:Interphase / Q000502:physiology / Q000528:radiation effects; D051379:Mice; D008938:Mitosis / Q000502:physiology / Q000528:radiation effects; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000502:physiology*
D016159:Tumor Suppressor Protein p53; D004247:DNA