Demonstration of pyrimidine dimer-DNA glycosylase activity in vivo: Bacteriophage T4-infected Escherichia coli as a model system

E. H. Radany, E. C. Friedberg

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Abstract

An approach to the detection of pyrimidine dimer-DNA glycosylate activity in living cells is presented. Mutants of Escherichia coli defective in uvr functions required for incision of UV-irradiated DNA were infected with phage T4 denV+ or denV- (defective in the T4 pyrimidine dimer-DNA glycosylate activity). In the former case the denV gene product catalyzed the incision of UV-irradiated host DNA, facilitating the subsequent excision of thymine-containing pyrimidine dimers. Isolation of these dimers from the acid-soluble fraction of infected cells was achieved by a multistep thin-layer chromatographic system. Exposure of the dimers to irradiation that monomerizes pyrimidine dimers (direct photoreversal) resulted in the stoichiometric formation of free thymine. Thus, in vivo incision of UV-irradiated DNA dependent on a pyrimidine dimer-DNA glycosylase can be demonstrated.

Original languageEnglish (US)
Pages (from-to)88-96
Number of pages9
JournalJournal of Virology
Volume41
Issue number1
Publication statusPublished - 1982

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ASJC Scopus subject areas

  • Immunology

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