Effects of p21(Cip1/Waf1) at both the G1/S and the G2/M cell cycle transitions: pRB is a critical determinant in blocking DNA replication and in preventing endoreduplication

Alexander B. Niculescu, Xinbin Chen, Monique Smeets, Ludger Hengst, Carol Prives, Steven I. Reed

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Abstract

It has been proposed that the functions of the cyclin-dependent kinase inhibitors p21(Cip1/Waf1) and p27(Kip1) are limited to cell cycle control at the G1/S-phase transition and in the maintenance of cellular quiescence. To test the validity of this hypothesis, p21 was expressed in a diverse panel of cell lines, thus isolating the effects of p21 activity from the pleiotropic effects of upstream signaling pathways that normally induce p21 expression. The data show that at physiological levels of accumulation, p21, in addition to its role in negatively regulating the G1/S transition, contributes to regulation of the G2/M transition. Both G1- and G2-arrested cells were observed in all cell types, with different preponderances. Preponderant G1 arrest in response to p21 expression correlated with the presence of functional pRb. G2 arrest was more prominent in pRb-negative cells. The arrest distribution did not correlate with the p53 status, and proliferating- cell nuclear antigen (PCNA) binding activity of p21 did not appear to be involved, since p27, which lacks a PCNA binding domain, produced similar arrest Bs. In addition, DNA endoreduplication occurred in pRb-negative but not in pRb-positive cells, suggesting that functional pRb is necessary to prevent DNA replication in p21 G2-arrested cells. These results suggest that the primary target of the Cip/Kip family of inhibitors leading to efficient G1 arrest as well as to blockade of DNA replication from either G1 or G2 phase is the pRb regulatory system. Finally, the tendency of Rb-negative cells to undergo endoreduplication cycles when p21 is expressed may have negative implications in the therapy of Rb-negative cancers with genotoxic agents that activate the p53/p21 pathway.

Original languageEnglish (US)
Pages (from-to)629-643
Number of pages15
JournalMolecular and cellular biology
Volume18
Issue number1
StatePublished - Jan 1 1998

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Endoreduplication
DNA Replication
Cell Cycle
Proliferating Cell Nuclear Antigen
G1 Phase
Cyclin-Dependent Kinase Inhibitor p21
G2 Phase
Phase Transition
Cell Cycle Checkpoints
S Phase
Maintenance
Cell Line
DNA

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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Effects of p21(Cip1/Waf1) at both the G1/S and the G2/M cell cycle transitions : pRB is a critical determinant in blocking DNA replication and in preventing endoreduplication. / Niculescu, Alexander B.; Chen, Xinbin; Smeets, Monique; Hengst, Ludger; Prives, Carol; Reed, Steven I.

In: Molecular and cellular biology, Vol. 18, No. 1, 01.01.1998, p. 629-643.

Research output: Contribution to journalArticle

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abstract = "It has been proposed that the functions of the cyclin-dependent kinase inhibitors p21(Cip1/Waf1) and p27(Kip1) are limited to cell cycle control at the G1/S-phase transition and in the maintenance of cellular quiescence. To test the validity of this hypothesis, p21 was expressed in a diverse panel of cell lines, thus isolating the effects of p21 activity from the pleiotropic effects of upstream signaling pathways that normally induce p21 expression. The data show that at physiological levels of accumulation, p21, in addition to its role in negatively regulating the G1/S transition, contributes to regulation of the G2/M transition. Both G1- and G2-arrested cells were observed in all cell types, with different preponderances. Preponderant G1 arrest in response to p21 expression correlated with the presence of functional pRb. G2 arrest was more prominent in pRb-negative cells. The arrest distribution did not correlate with the p53 status, and proliferating- cell nuclear antigen (PCNA) binding activity of p21 did not appear to be involved, since p27, which lacks a PCNA binding domain, produced similar arrest Bs. In addition, DNA endoreduplication occurred in pRb-negative but not in pRb-positive cells, suggesting that functional pRb is necessary to prevent DNA replication in p21 G2-arrested cells. These results suggest that the primary target of the Cip/Kip family of inhibitors leading to efficient G1 arrest as well as to blockade of DNA replication from either G1 or G2 phase is the pRb regulatory system. Finally, the tendency of Rb-negative cells to undergo endoreduplication cycles when p21 is expressed may have negative implications in the therapy of Rb-negative cancers with genotoxic agents that activate the p53/p21 pathway.",
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