A role for p53 in maintaining and establishing the quiescence growth arrest in human cells.
The p53 tumor suppressor protein induces transient growth arrest or apoptosis in response to genotoxic stress and mediates the irreversible growth arrest of cellular senescence. We present evidence here that p53 also contributes to the reversible, growth factor-dependent arrest of quiescence (G(0)). Microinjection of expression vectors encoding either MDM2 or a pRb-binding mutant of SV40 T antigen, both of which abrogate p53 function, stimulated quiescent normal human fibroblasts to initiate DNA synthesis and were 40-70% as effective as wild-type T antigen. Electrophoretic mobility shift and p53 transactivation assays showed that p53 activity was higher in quiescent and senescent cells compared with proliferating cells. As proliferating cells entered G(0) after growth factor withdrawal, the p53 mRNA level increased, followed by transient accumulation of the protein. Shortly thereafter, the expression (mRNA and protein) of p21, a p53 target gene and effector of cell cycle arrest, increased. Finally, stable expression of the HPV16 E6 oncogene or dominant negative p53 peptide, GSE-22, both of which inhibit p53 function, delayed entry into quiescence following growth factor withdrawal. Our data indicate that p53 is activated during both quiescence and senescence. They further suggest that p53 activity contributes, albeit not exclusively, to the quiescent growth arrest.
Itahana
Koji
K
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Dimri
Goberdhan P
GP
Hara
Eiji
E
Itahana
Yoko
Y
Zou
Ying
Y
Desprez
Pierre-Yves
PY
Campisi
Judith
J
D000957:Antigens, Viral, Tumor / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology*; D002478:Cells, Cultured; D004261:DNA Replication; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D008845:Microinjections; D012333:RNA, Messenger / Q000378:metabolism; D016192:Resting Phase, Cell Cycle; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
D000957:Antigens, Viral, Tumor; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53