Use of shuttle vectors to study the molecular processing of defined carcinogen-induced DNA damage: mutagenicity of single O4-ethylthymine adducts in HeLa cells
| Autor: | J.T. Lutgerink, J.G. Westra, E. Kriek, M.J. BIeeker, W.J. van Dijk, J. H. Van Boom, H. F. Brugghe, H.den van Elst, J. C. Klein, A.J.M. Bems, G.A.der van Marel |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
DNA Replication
DNA polymerase DNA damage Genetic Vectors Molecular Sequence Data Transfection Cell Line chemistry.chemical_compound Shuttle vector Genetics Humans Base Sequence biology Mutagenicity Tests Oligonucleotide Mutagenesis DNA replication Molecular biology Kinetics Oligodeoxyribonucleotides chemistry Adductomics Biochemistry Mutation Carcinogens biology.protein Thymine DNA DNA Damage HeLa Cells |
| Zdroj: | Nucleic Acids Research. 18:4131-4137 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/18.14.4131 |
| Popis: | We developed a simian virus 40 based shuttle vector system to study the molecular consequences of distinct carcinogen-induced DNA lesions in human cells. To establish the mutagenicity of O4-ethylthymine adducts, oligonucleotides carrying a single O4-ethylthymine adduct at a unique position were ligated into the vector molecules. Following replication in HeLa cells on average 23% of the progeny molecules carried a mutation in the region of modification. The vast majority of these mutations represented single T----C transitions at the position of the modified base, most probably as a consequence of mispairing of the O4-ethylthymine residues during replication. To a minor extent the O4-ethylthymine adduct may also induce T----A transversions or double point mutations. The in vivo mutation frequency of the adduct was found to be comparable to that of a C-A mismatch at the same position, but was lower than that expected from in vitro experiments with adducted DNA templates and purified DNA polymerases. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|