Cell cycle dependent effects of u.v.-radiation on p53 expression and retinoblastoma protein phosphorylation.
Control of fate of cells encountered with DNA damaging agents is pivotal for normal cellular homeostasis. DNA damage leads in many cases to growth arrest of the cells ensuring sufficient time for damage repair. Growth arrest can be mediated by p53 tumor suppressor protein and loss of its function leads to inability of the cells to both growth arrest and undergo apoptosis. We show here that followed by genotoxic stress, the retinoblastoma gene product, pRB, is associated with growth arrest of cells in a p53 independent manner. In u.v.-treated human and mouse fibroblasts, pRB is rapidly dephosphorylated. pRB dephosphorylation occurs concomitant with growth arrest of cells including cells with p53 mutations (SW 480 colon carcinoma cells), cells expressing SV40 T antigen and rat-transformed cells (T-24 bladder carcinoma cells) unresponsive in regard to p53 stimulation. Furthermore, flow cytometry analysis of u.v.-radiated synchronized G1 cells indicates that the cells transiently arrest in G1 for 10-12 h with pRB dominating in its underphosphorylated form, whereas p53 accumulation occurs only after the cells have entered into S-phase. In addition, u.v.-radiation of late S- and G2/M-phase cells leads to p53 accumulation and cell cycle arrest. The results indicate that p53 accumulation upon u.v.-radiation occurs during DNA replication and is thus not involved in G1 arrest. We suggest that the events that lead to pRB dephosphorylation upon u.v.-radiation provide the cell an efficient G1 arrest which occurs prior and independently of p53.
Haapajärvi
T
T
Department of Virology, University of Helsinki, Finland.
Kivinen
L
L
Pitkänen
K
K
Laiho
M
M
D016475:3T3 Cells; D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002460:Cell Line; D004261:DNA Replication / Q000528:radiation effects; D006801:Humans; D051379:Mice; D010766:Phosphorylation; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D014466:Ultraviolet Rays
D016160:Retinoblastoma Protein; D016159:Tumor Suppressor Protein p53