NDLI: p53-dependent growth arrest of REF52 cells containing newly amplified DNA.

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p53-dependent growth arrest of REF52 cells containing newly amplified DNA.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Ishizaka, Y. Chernov, M. V. Burns, C. M. Stark, G. R.
Description	Author Affiliation: Ishizaka Y ( Department of Molecular Biology, Cleveland Clinic Foundation, OH 44195, USA.);
Abstract	The rat cell line REF52 is not permissive for gene amplification. Simian virus 40 tumor (T) antigen converts these cells to a permissive state, as do dominant negative mutants of p53, suggesting that the effect of T antigen is due mainly to its ability to bind to p53. To manipulate permissivity, we introduced a temperature-sensitive mutant of T antigen (tsA58) into REF52 cells and selected for resistance to N-(phosphonacetyl)-L-aspartate (PALA). Most freshly isolated PALA-resistant colonies, each of approximately 200 cells, selected at a permissive temperature, arrested when shifted to a nonpermissive temperature. Growth arrest was stable, with no evidence of apoptosis, as long as T antigen was absent but was reversed when T antigen was restored. In contrast, PALA-resistant clones grown to approximately 10(7) cells at a permissive temperature did not arrest when shifted to a nonpermissive temperature. All PALA-resistant clones examined had amplified carbamoyl-phosphate synthetase-aspartate transcarbamoylase-dihydroorotase (CAD) genes, present in structures consistent with a mechanism involving bridge-breakage-fusion (BBF) cycles. We propose that p53-mediated growth arrest operates only early during the complex process of gene amplification, when newly formed PALA-resistant cells contain broken DNA, generated in BBF cycles. During propagation under permissive conditions, the broken DNA ends are healed, and, even though the p53-mediated pathway is still intact at a nonpermissive temperature and the cells contain amplified DNA, they are not arrested in the absence of broken DNA. The data support the hypothesis that BBF cycles are an important mechanism of amplification and that the broken DNA generated in each cycle is a key signal that regulates permissivity for gene amplification.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	8
Volume Number	92
Language	English
Publisher	National Academy of Sciences
Publisher Date	1995-05-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Antigens, Viral, Tumor Physiology Cell Cycle Gene Amplification Tumor Suppressor Protein P53 Animals Genetics Aspartate Carbamoyltransferase Aspartic Acid Analogs & Derivatives Pharmacology Carbamyl Phosphate Metabolism Cell Line Cyclin-Dependent Kinase Inhibitor P21 Cyclins DNA Damage DNA Replication Dihydroorotase Drug Resistance Hot Temperature In Situ Hybridization, Fluorescence Ligases Mutation Phosphonoacetic Acid RNA, Messenger Biosynthesis Recombinant Proteins Selection, Genetic Simian Virus 40 Research Support, Non-U.S. Gov't Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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