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Cell-free assay measuring repair DNA synthesis in human fibroblasts

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
;
Osmotic disruption of confluent cultured human fibroblasts that have been irradiated or exposed to chemical carcinogens allows the specific measurement of repair DNA synthesis using dTTP as a precursor. Fibroblasts similarly prepared from various xeroderma pigmentosum cell lines show the deficiencies of uv-induced DNA synthesis predicted from in vivo studies, while giving normal responses to methylmethanesulfonate. A pyrimidine-dimer-specific enzyme, T4 endonuclease V, stimulated the rate of uv-induced repair synthesis with normal and xeroderma pigmentosum cell lines. This system should prove useful for identifying agents that induce DNA repair, and cells that respond abnormally to such induction. It should also be applicable to an in vitro complementation assay with repair-defective cells and proteins obtained from repair-proficient cells. Finally, by using actively growing fibroblasts and thymidine in the system, DNA replication can be measured and studied in vitro.
Research Organization:
Univ. of California, Berkeley
OSTI ID:
6848286
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 75:4; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

560121* -- Radiation Effects on Cells-- External Source-- (-1987)
560301 -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CARCINOGENS
CELL CULTURES
COMPARATIVE EVALUATIONS
CONNECTIVE TISSUE CELLS
DNA
DNA REPLICATION
ELECTROMAGNETIC RADIATION
FIBROBLASTS
IONIZING RADIATIONS
IRRADIATION
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIATION EFFECTS
RADIATIONS
RADIOINDUCTION
RECOVERY
REPAIR
SOMATIC CELLS
ULTRAVIOLET RADIATION
X RADIATION
XP CELLS