Repair of site-specific double-strand breaks in a mammalian chromosome by homologous and illegitimate recombination
| Autor: | R G Sargent, John H. Wilson, Mark A. Brenneman |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Mitotic crossover
Saccharomyces cerevisiae Proteins DNA Repair DNA repair FLP-FRT recombination Molecular Sequence Data Non-allelic homologous recombination Adenine Phosphoribosyltransferase CHO Cells Saccharomyces cerevisiae Biology Genetic recombination Chromosomes Cricetinae Animals Point Mutation Ectopic recombination Deoxyribonucleases Type II Site-Specific Molecular Biology Genetics Gene Rearrangement Recombination Genetic Base Sequence Cell Biology Sequence Analysis DNA Non-homologous end joining Gene Targeting Homologous recombination DNA Damage Research Article |
| Zdroj: | Molecular and cellular biology. 17(1) |
| ISSN: | 0270-7306 |
| Popis: | In mammalian cells, chromosomal double-strand breaks are efficiently repaired, yet little is known about the relative contributions of homologous recombination and illegitimate recombination in the repair process. In this study, we used a loss-of-function assay to assess the repair of double-strand breaks by homologous and illegitimate recombination. We have used a hamster cell line engineered by gene targeting to contain a tandem duplication of the native adenine phosphoribosyltransferase (APRT) gene with an I-SceI recognition site in the otherwise wild-type APRT+ copy of the gene. Site-specific double-strand breaks were induced by intracellular expression of I-SceI, a rare-cutting endonuclease from the yeast Saccharomyces cerevisiae. I-SceI cleavage stimulated homologous recombination about 100-fold; however, illegitimate recombination was stimulated more than 1,000-fold. These results suggest that illegitimate recombination is an important competing pathway with homologous recombination for chromosomal double-strand break repair in mammalian cells. |
| Databáze: | OpenAIRE |
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