Loss-of-function genetics in mammalian cells: The p53 tumor suppressor model

Carnero, A., Hudson, J. D., Hannon, G. J., Beach, D. H. (2000) Loss-of-function genetics in mammalian cells: The p53 tumor suppressor model. Nucleic Acids Research, 28 (11). pp. 2234-2241. ISSN 03051048 (ISSN)

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URL: http://www.ncbi.nlm.nih.gov/pubmed/10871344
DOI: 10.1093/nar/28.11.2234

Abstract

Using an improved system for the functional identification of active antisense fragments, we have isolated antisense fragments which inactivate the p53 tumour suppressor gene. These antisense fragments map in two small regions between nt 350 and 700 and nt 800 and 950 of the coding sequence. These antisense fragments appear to act by inhibition of p53 mRNA translation both in vivo and in vitro. Expression of these antisense fragments overcame the p53-induced growth arrest in a cell line which expresses a thermolabile mutant of p53 and extended the in vitro lifespan of primary mouse embryonic fibroblasts. Continued expression of the p53 antisense fragment contributed to immortalisation of primary mouse fibroblasts. Subsequent elimination of the antisense fragment in these immortalised cells led to restoration of p53 expression and growth arrest, indicating that immortal cells continuously require inactivation of p53. Expression of MDM2 or SV40 large T antigen, but not E7 nor oncogenic ras, overcomes the arrest induced by restoration of p53 expression. Functional inactivation of both p21 and bax (by overexpression of Bcl2), but not either alone, allowed some bypass of p53-induced growth arrest, indicating that multiple transcriptional targets of p53 may mediate its antiproliferative action. The ability to conditionally inactivate and subsequently restore normal gene function may be extremely valuable for genetic analysis of genes for which loss-of-function is involved in specific phenotypes.

Item Type: Paper
Uncontrolled Keywords: apolipoprotein E7 messenger RNA mutant protein protein Bax protein bcl 2 protein MDM2 protein p21 protein p53 virus T antigen animal cell article cell growth cell proliferation controlled study embryo embryo cell fibroblast gene function gene overexpression gene sequence genetic analysis genetic code growth inhibition in vitro study in vivo study lifespan mammal cell mouse nonhuman nucleic acid probe oncogene ras phenotype priority journal protein expression protein targeting RNA translation Simian virus 40 somatic cell genetics translation regulation tumor suppressor gene Animals Antigens, Viral, Tumor Fibroblasts Gene Expression Regulation Mice Mutation Nuclear Proteins Protein Biosynthesis Proto-Oncogene Proteins Proto-Oncogene Proteins c-bcl-2 Proto-Oncogene Proteins c-mdm2 RNA, Antisense RNA, Messenger Tumor Suppressor Protein p53 Animalia Mammalia Simiae Simian virus
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > loss of function
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > p53
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > suppressor
CSHL Authors:
Communities: CSHL labs > Hannon lab
Depositing User: Matt Covey
Date: 2000
Date Deposited: 09 Jan 2013 17:33
Last Modified: 09 Jan 2013 17:33
PMCID: PMC102629
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26477

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