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Mol Cell Biol, February 1998, p. 1055-1064, Vol. 18, No. 2
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Characterization of the p53-Dependent Postmitotic Checkpoint following Spindle Disruption

Jennifer S. Lanni1,2 and Tyler Jacks1,2,3,*

Center for Cancer Research,1 Department of Biology,2 and Howard Hughes Medical Institute,3 Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Received 5 September 1997/Returned for modification 9 October 1997/Accepted 10 November 1997

The p53 tumor suppressor gene product is known to act as part of a cell cycle checkpoint in G1 following DNA damage. In order to investigate a proposed novel role for p53 as a checkpoint at mitosis following disruption of the mitotic spindle, we have used time-lapse videomicroscopy to show that both p53+/+ and p53-/- murine fibroblasts treated with the spindle drug nocodazole undergo transient arrest at mitosis for the same length of time. Thus, p53 does not participate in checkpoint function at mitosis. However, p53 does play a critical role in nocodazole-treated cells which have exited mitotic arrest without undergoing cytokinesis and have thereby adapted. We have determined that in nocodazole-treated, adapted cells, p53 is required during a specific time window to prevent cells from reentering the cell cycle and initiating another round of DNA synthesis. Despite having 4N DNA content, adapted cells are similar to G1 cells in that they have upregulated cyclin E expression and hypophosphorylated Rb protein. The mechanism of the p53-dependent arrest in nocodazole-treated adapted cells requires the cyclin-dependent kinase inhibitor p21, as p21-/- fibroblasts fail to arrest in response to nocodazole treatment and become polyploid. Moreover, p21 is required to a similar extent to maintain cell cycle arrest after either nocodazole treatment or irradiation. Thus, the p53-dependent checkpoint following spindle disruption functionally overlaps with the p53-dependent checkpoint following DNA damage.

* Corresponding author. Mailing address: MIT Center for Cancer Research, 40 Ames St., Building E17-517, Cambridge, MA 02139. Phone: (617) 253-0262. Fax: (617) 253-9863. E-mail: tjacks{at}mit.edu.




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