Gene activation by induced DNA rearrangements

Schnipper, L E; Chan, V; Sedivy, J; Jat, P; Sharp, P A

Title: Gene activation by induced DNA rearrangements

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Abstract

A murine cell line (EN/NIH) containing the retroviral vector ZIPNeoSV(x)1 that was modified by deletion of the enhancer elements in the viral long terminal repeats has been used as an assay system to detect induced DNA rearrangements that result in activation of a transcriptionally silent reporter gene encoded by the viral genome. The spontaneous frequency of G418 resistance is less than 10(-7), whereas exposure to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) or the combination of UV irradiation plus TPA resulted in the emergence of drug resistant cell lines at a frequency of 5 per 10(6) and 67 per 10(6) cells, respectively. In several of the cell lines that were analyzed a low level of amplification of one of the two parental retroviral integrants was observed, whereas in others no alteration in the region of the viral genome was detected. To determine the effect of the SV40 large T antigen on induced DNA rearrangements, EN/NIH cells were transfected with a temperature sensitive (ts) mutant of SV40 T. Transfectants were maintained at the permissive temperature (33 degrees C) for varying periods of time (1-5 days) in order to vary SV40 T antigen exposure, after which they were shifted to 39.5 degrees C formore »« less

Authors:

Schnipper, L E; Chan, V; Sedivy, J; Jat, P; Sharp, P A ^[1]

Thorndike Memorial Laboratories, Dana Research Institute, Boston, MA (USA)

Publication Date:: Fri Dec 01 00:00:00 EST 1989

OSTI Identifier:: 7164918

Resource Type:: Journal Article

Journal Name:: Cancer Research; (USA)

Additional Journal Information:: Journal Volume: 49:23; Journal ID: ISSN 0008-5472

Country of Publication:: United States

Language:: English

Subject:: 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; GENE MUTATIONS; RADIOINDUCTION; PHORBOL ESTERS; GENETIC EFFECTS; ANTIGENS; GENE AMPLIFICATION; GENE RECOMBINATION; GENE REGULATION; GENETIC RADIATION EFFECTS; MICE; MUTATION FREQUENCY; PHOSPHOTRANSFERASES; ULTRAVIOLET RADIATION; VIRUSES; ANIMALS; BIOLOGICAL EFFECTS; BIOLOGICAL RADIATION EFFECTS; CARCINOGENS; ELECTROMAGNETIC RADIATION; ENZYMES; ESTERS; MAMMALS; MICROORGANISMS; MUTATIONS; ORGANIC COMPOUNDS; PARASITES; PHOSPHORUS-GROUP TRANSFERASES; RADIATION EFFECTS; RADIATIONS; RODENTS; TRANSFERASES; VERTEBRATES; 560120* - Radiation Effects on Biochemicals, Cells, & Tissue Culture; 560300 - Chemicals Metabolism & Toxicology

Citation Formats


                    Schnipper, L E, Chan, V, Sedivy, J, Jat, P, and Sharp, P A. Gene activation by induced DNA rearrangements.  United States: N. p., 1989. 
        Web.

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                    Schnipper, L E, Chan, V, Sedivy, J, Jat, P, & Sharp, P A. Gene activation by induced DNA rearrangements.  United States.

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                    Schnipper, L E, Chan, V, Sedivy, J, Jat, P, and Sharp, P A. 1989.  
        "Gene activation by induced DNA rearrangements".  United States.

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@article{osti_7164918,

  title        = {Gene activation by induced DNA rearrangements},

  author       = {Schnipper, L E and Chan, V and Sedivy, J and Jat, P and Sharp, P A},

  abstractNote = {A murine cell line (EN/NIH) containing the retroviral vector ZIPNeoSV(x)1 that was modified by deletion of the enhancer elements in the viral long terminal repeats has been used as an assay system to detect induced DNA rearrangements that result in activation of a transcriptionally silent reporter gene encoded by the viral genome. The spontaneous frequency of G418 resistance is less than 10(-7), whereas exposure to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) or the combination of UV irradiation plus TPA resulted in the emergence of drug resistant cell lines at a frequency of 5 per 10(6) and 67 per 10(6) cells, respectively. In several of the cell lines that were analyzed a low level of amplification of one of the two parental retroviral integrants was observed, whereas in others no alteration in the region of the viral genome was detected. To determine the effect of the SV40 large T antigen on induced DNA rearrangements, EN/NIH cells were transfected with a temperature sensitive (ts) mutant of SV40 T. Transfectants were maintained at the permissive temperature (33 degrees C) for varying periods of time (1-5 days) in order to vary SV40 T antigen exposure, after which they were shifted to 39.5 degrees C for selection in G418. The frequency of emergence of drug resistant cell clones increased with duration of exposure to large T antigen (9-52 per 10(6) cells over 1-5 days, respectively), and all cell lines analyzed demonstrated DNA rearrangements in the region of the neo gene. A novel 18-kilobase pair XbaI fragment was cloned from one cell line which revealed the presence of a 2.0-kilobase pair EcoRI segment containing an inverted duplication which hybridized to neo sequences. It is likely that the observed rearrangement was initiated by the specific binding of large T antigen to the SV40 origin of replication encoded within the viral genome.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/7164918},
  journal      = {Cancer Research; (USA)},

  issn         = {0008-5472},

  number       = ,

  volume       = 49:23,

  place        = {United States},

  year         = {Fri Dec 01 00:00:00 EST 1989},

  month        = {Fri Dec 01 00:00:00 EST 1989}

}

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