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

Journal Article · Fri Jan 01 04:00:00 EST 1982 · J. Virol.; (United States)

OSTI ID:6723184

Radany, E H; Friedberg, E C

An approach to the detection of pyrimidine dimer-DNA glycosylase 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 glycosylase 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.

• Research Organization: Department of Pathology, Stanford University, CA
• OSTI ID: 6723184
• Journal Information: J. Virol.; (United States), Journal Name: J. Virol.; (United States) Vol. 41:1; ISSN JOVIA
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
550700 -- Microbiology
560121* -- Radiation Effects on Cells-- External Source-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AZINES
BACTERIA
BACTERIOPHAGES
BIOCHEMISTRY
BIOLOGICAL EFFECTS
BIOLOGICAL PATHWAYS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CHEMISTRY
DIMERS
DNA
ELECTROMAGNETIC RADIATION
ENZYME ACTIVITY
ENZYMES
ESCHERICHIA COLI
GLYCOSYL HYDROLASES
HETEROCYCLIC COMPOUNDS
HYDROLASES
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PYRIMIDINES
RADIATION EFFECTS
RADIATIONS
RECOVERY
REPAIR
ULTRAVIOLET RADIATION
VIRUSES