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 language | English (US) |
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Pages (from-to) | 88-96 |
Number of pages | 9 |
Journal | Journal of virology |
Volume | 41 |
Issue number | 1 |
DOIs | |
State | Published - 1982 |
ASJC Scopus subject areas
- Microbiology
- Immunology
- Insect Science
- Virology